Endocrine disruption in soil invertebrates

Sophie Campiche
EPFL, 2006
EPFL thesis

Abstract

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

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Sophie Campiche
EPFL, 2006
EPFL thesis

Abstract

Official source

https://infoscience.epfl.ch/record/64689?ln=en

About this result

Related concepts (43)

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Due to human activities, organic pollutants are spilled to the environment where they threaten public health, often as contaminants of soil or groundwater. Living organisms are able to transform or mineralize many organic pollutants, and bioremediation techniques have been developed to remove pollutants from a contaminated site. However, fast and easy methods to document both the efficacy of bioremediation and the changes in soil microbial communities during bioremediation are not well developed. The major aim of this thesis was to develop miniaturized methods targeting the physiology of microbial communities during pollutant degradation in soil and to assess pollution-induced community changes. Furthermore, the methods should identify factors limiting efficient pollutant biodegradation. Petroleum hydrocarbons have been chosen as pollutants because these compounds are frequently spilled, readily water-soluble, partly toxic and carcinogenic, and therefore undesirable in soils and drinking water supplies. The influence of petroleum hydrocarbons on a microbial community in the vadose zone was assessed under field conditions. An artificial hydrocarbon mixture consisting of volatile and semi-volatile compounds similar to jet-fuel was emplaced in a previously uncontaminated vadose zone in nutrient-poor glacial melt water sand. The experiment included monitoring of microbial parameters and CO2 concentrations in soil gas over 3 months, in- and outside the hydrocarbon vapor plume that formed around the buried petroleum. Microbial and chemical analyses of vadose zone samples were performed on material from 9 cores drilled on 3 days at 3 distances from the buried petroleum mass to 3.3 m depth. Significantly elevated CO2 concentrations were observed after contamination. Total cell numbers as determined by fluorescence microscopy were strongly correlated with soil organic carbon and nitrogen content but varied little with contamination. Redundancy analysis (RDA) allowed direct analysis of effects of selected environmental variables or the artificial contamination on microbiological parameters. Variation in biomass and CO2 production was to 46 % explained by soil parameters and to 39.8 % by the duration of contamination. The microbial community structure was assessed by community-level physiological profile analysis using BiologTM EcoPlates. Only 35.9 % of variation in BiologTM data could be attributed to soil parameters and contamination, however, the samples with greatest exposure to hydrocarbons grouped together on RDA plots. It is concluded that at this nutrient-poor site, the microbial community was dominated by the natural heterogeneity and that the influence of petroleum hydrocarbon vapor was weak. BiologTM Eco-Plates combined with RDA were able to distinguish between samples with high and low hydrocarbon exposure. However, the method was not sensitive enough to produce consistent patterns in three replicates when extracts from subsurface soils with low cell numbers were inoculated. A more sensitive, MPN-based system was therefore developed and its ability to provide information about specific pollutant degraders was tested in alluvial sand. Eight dilutions of sand extracts were inoculated on medium- and substrate-containing polypropylene deep-well plates. To keep the concentrations of the volatile substrates constant over several days of incubation without intoxicating the soil community, a substrate-containing organic carrier phase was applied to each vial. The biochemically inert 2,2,4,4,6,8,8-heptamethylnonane (HMN) was appropriate for this purpose. Thus, the volatile substrates partition from HMN into the microorganism-containing soil water where they achieve relatively low concentrations. The sensitive watersoluble tetrazolium dye (WST-1) was added to each well after 7 days incubation to detect dehydrogenase activity. A dark yellow signal was developed for the substrates toluene, n-octane, n-dodecane, 1,2,4-trimethylbenzene and methylcyclohexane but not for isooctane, being strongest in n-alkane amended wells. With this method, more hydrocarbon degrading bacteria were detected in an alluvial sand exposed to kerosene for 72 hours compared to the community in the pristine sand. Both communities were mainly composed of n-alkane degraders. Although encouraging results were achieved, we noticed that the tetrazolium reduction was inhibited in some cases. Wells with high cell numbers, in which obvious growth occurred, did not produce a signal. In a next step, the two-liquid phase system with HMN was applied to modify a miniaturized respiration detection system using whole soil samples instead of soil extracts (Campbell et al., AEM 69, p. 3593). Soil was incubated in deep-well plates in presence of single volatile organic hydrocarbons. The carrier phase was added to the interstitial space in a bead layer on the bottom of the well (below the soil sample), and the hydrocarbons diffused into the soil-water. The soil activity was determined by means of an agar plate containing a pH indicator, which changes color as a function of the produced CO2. Physiological profiles specific to petroleum hydrocarbons in pristine and contaminated soils were assessed. The substrate concentrations, which induced highest microbial activity, as well as concentrations causing inhibition, were determined with effect-concentration curves. Community-level physiological profiles based on hydrocarbon degradation were obtained by applying multivariate analysis on the CO2 yields. The first measurement period of 0-6 hours after addition of relatively high hydrocarbon concentrations to the system yields values, which best separated soil types as well as pristine and contaminated soils. If low concentrations were applied, we obtained only significant separations after 24 hours incubation, probably after a growth step. The same micro-respiration system equipped with a 14CO2 detection plate was applied to study N and P limitation of aerobic mineralization of 14C-toluene in a soil previously exposed to petroleum. Significant nutrient limitation during short-term tests (24-48 hours) was identified and the optimized combination of nutrient addition was determined. Conclusively, miniaturized physiological methods have been proven to detect soil microbial community changes in petroleum hydrocarbon-contaminated soils. Adding an organic carrier phase to the miniaturized systems allowed assuring a volatile organic hydrocarbon supply in non-toxic concentrations in the soil water or the medium for several days. Thus simple, cost-effective systems to investigate the soil community during bioremediation and to address questions of nutrient shortage in soils have been provided with this thesis.

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

Organic pollutants in Swiss compost and digestate

Rahel Christine Brändli

Composting (aerobic treatment of organic wastes) and digestion (anaerobic treatment of organic wastes combined with biogas production) are important waste management strategies with increasing significance in the European Union and Switzerland. Most of the compost produced is applied to agricultural soils, which hereby recycles nutrients and influences soil properties beneficially. However, compost can contain pollutants that may be hazardous for the soil ecosystem. The problem related to heavy metals had been recognised and measures for reduction were taken. Regarding organic pollutants, the current knowledge is insufficient for quality control and risk assessment. This thesis provides a comprehensive overview on organic contaminates in compost, digestate, and presswater and describes factors that may influence them. In the beginning an extensive literature review was carried out to summarize the current data. Compound classes to be analysed in Swiss composts and digestates were prioritized and analytical methods established. Polychlorinated biphenyl concentrations (∑ of PCB 28, 52, 101, 118, 138, 153, 180) were significantly higher in urban (median: 30 μg/kgdry weight (dw), n=52) than in rural samples (median: 14 μg/kgdw, n=16), which points – together with low concentrations in general – to aerial deposition on feedstock material as major input pathway to compost. Median polycyclic aromatic hydrocarbon (PAH) concentration was 3010 μg/kgdw (∑ of 16 PAH defined by the US EPA except dibenzo[a,h]anthracene, n=69). PAH levels were influenced by the organic matter degradation process (higher levels in digestate than in compost), the season of input material collection (spring/summer>winter>autumn), the particle size (higher concentrations in unsieved or sieved >20mm than in sieved to ≤ 20mm products), and maturity (lower concentrations in more mature composts). One fourth of the samples exhibited PAH concentrations above the Swiss guide value for compost (Ordinance on the Reductions of Risks linked to Chemical Products). These elevated concentrations can lead to considerable input of PAH to soil by compost application. To assess the major contributors of PAH in compost, characteristic PAH ratios and some molecular markers were considered, which pointed mainly to combustion origin of these contaminants. Multifactor statistical analysis indicated traffic emission, straw combustion and some asphalt abrasion as potential additional sources. Concentrations of other organic pollutants determined (dibenzo-p-dioxins and -furans, dioxin-like PCB, brominated flame retardants (BFR), perfluorinated alkyl substances (PFAS), di(2-ethylhexyl)phthalate, nonylphenol and chlorinated paraffins) were mostly above levels found in background soil, except for nonylphenol, which was not detected. Out of 269 pesticides analysed, 30 fungicides, 14 herbicides, eight insecticides and one acaricide were detected. For the first time, the fate of organic pollutants during full-scale composting and digestion was assessed. Concentrations of low chlorinated PCB increased during composting (about 30 %), whereas a slight decrease was observed for the higher chlorinated congeners (about 10%). Enantiomeric ratios of atropisomeric PCB were close to racemic and did not change. Levels of low molecular weight PAH were reduced during composting (50 to 90% reduction), whereas heavier compounds remained stable. However, as indicated above, conventional composting does not reduce PAH concentrations sufficiently to comply with Swiss guide values (see above). Further research is needed to i) identify measures to reduce PAH concentrations in digestate and compost, ii) to monitor organic pollutants that are still increasing in other environmental matrices (e.g. BFR and PFAS) or identify new compounds which have not been detected in compost yet, iii) to evaluate potential risk of compost application to soil by assessing the bioavailable fractions of organic pollutants and iv) to examine possible new input materials and co-substrates for composting and digestion.

EPFL2006

Organic pollutants in Swiss compost and digestate

Rahel Christine Brändli

EPFL2006

EPFL2004

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

Jérôme Payet

EPFL2004