Investigating predictive tools for refinery effluent hazard assessment using stream mesocosms.

Hazard assessment of refinery effluents is challenging because of their compositional complexity. Therefore, a weight-of-evidence approach using a combination of tools is often required. Previous research has focused on several predictive tools for sophisticated chemical analyses: biomimetic extraction to quantify the potentially bioaccumulative substances, 2-dimensional gas chromatography, modeling approaches to link oil composition to toxicity (PETROTOX), and whole-effluent toxicity assessments using bioassays. The present study investigated the value of these tools by comparing predicted effects to actual effects observed in stream mesocosm toxicity studies with refinery effluents. Three different effluent samples, with and without fortification by neat petroleum substances, were tested in experimental freshwater streams. The results indicate that the biological community shifted at higher exposure levels, consistent with chronic toxicity effects predicted by both modeled toxic units and potentially bioaccumulative substance measurements. The present study has demonstrated the potential of the predictive tools and the robustness of the stream mesocosm design to improve our understanding of the environmental hazards posed by refinery effluents. Environ Toxicol Chem 2019;38:650-659. © 2018 SETAC.

Sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants.

Hydrocarbons have an utmost economical importance but may also cause substantial ecological impacts due to accidents or inadequate transportation and use. Currently, freshwater biomonitoring methods lack an indicator that can unequivocally reflect the impacts caused by hydrocarbons while being independent from effects of other stressors. The aim of the present study was to develop a sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants, which can be used in hydrocarbon-specific bioindicators. We employed the Relative Sensitivity method and developed the sensitivity ranking S hydrocarbons based on literature ecotoxicological data supplemented with rapid and mesocosm test results. A first validation of the sensitivity ranking based on an earlier field study has been conducted and revealed the S hydrocarbons ranking to be promising for application in sensitivity based indicators. Thus, the first results indicate that the ranking can serve as the core component of future hydrocarbon-specific and sensitivity trait based bioindicators.

Differential protein expression in the estuarine copepod Eurytemora affinis after diuron and alkylphenol exposures.

Proteomics was used in the calanoid copepod Eurytemora affinis for screening of protein expression modifications induced by organic contaminants. The copepods were exposed in a continuous flow-through system for 86 h to environmentally relevant concentrations of contaminants representative of the pollution in the Seine Estuary (Haute-Normandie, France; diuron, 500 ng L(-1) ; alkylphenol mixture, 1000 ng L(-1) ). Proteome analysis of whole-body copepod extracts by 2-dimensional gel electrophoresis revealed that the contaminants induced modifications in protein expression, with the highest quantitative variations occurring after diuron exposure. Specifically, 88 and 41 proteins were differentially expressed after diuron and alkylphenol treatments, respectively. After mass spectrometry analysis, 51 (diuron exposure) and 15 (alkylphenol exposure) proteins were identified. The identified proteins were potentially related to energy metabolism, cell growth, nervous signal conductivity, excitotoxicity, oxidative stress response, and antioxidant defense. The data suggest a massive general disturbance of physiological functions of E. affinis after diuron exposure, whereas alkylphenols induced an alteration of a few targeted physiological functions. The protein expression signatures identified after contaminant exposure deserve further investigation in terms of the development of novel potential biomarkers for water quality assessment. Environ Toxicol Chem 2016;35:1860-1871. © 2015 SETAC.

Effect of thiram and of a hydrocarbon mixture on freshwater macroinvertebrate communities in outdoor stream and pond mesocosms: I. Study design, chemicals fate and structural responses.

Higher-tier ecological risk assessment (ERA) in mesocosms is commonly performed in lotic or lentic experimental systems. These systems differ in their physico-chemical and hydrological properties, leading to differences in chemical fate, community characteristics and potential recovery. This raises the issue of the relevance and sensitivity of community-level endpoints in different types of mesocosms. In this study, macroinvertebrate abundance and biomass estimates were used to assess the effects of a dithiocarbamate fungicide, thiram (35 and 170 µg l(-1)), and a petroleum middle distillate (PMD; 0.01, 0.4, 2 and 20 mg l(-1)) in outdoor stream and pond mesocosms. Streams were continuously treated during 3 weeks followed by a 2-month long post-treatment period. Ponds were treated weekly for 4 weeks, followed by a 10-month long post-treatment period. Taxonomic structure of macroinvertebrate communities was characterized using the α, ß and γ components of taxa richness, Shannon and Gini-Simpson indices. Computations were based either on abundance or biomass data. Results clearly highlighted that the effects of chemicals depended on the exposure regime (for thiram) and type of system (for the PMD). Causes of the differences between streams and ponds in the magnitude and nature of effects include differential sensitivity of taxa dwelling in lentic and lotic systems and the influence of hydrology (e.g., drift from upstream) and mesocosm connectivity on recovery dynamics. This study also showed complementarities in the use of both types of mesocosms to improve the characterization of chemical effects on communities in ERA.

Effect of thiram and of a hydrocarbon mixture on freshwater macroinvertebrate communities in outdoor stream and pond mesocosms: II. Biological and ecological trait responses and leaf litter breakdown.

Higher-tier ecological risk assessment of chemicals often relies upon studies in dynamic and/or static mesocosms. Physico-chemical and hydrological properties of each type of mesocosm result in specific chemicals fate, community functioning, and potential recovery. In the present study, macroinvertebrate abundance- and biomass-weighted biological and ecological trait matrices were used to assess the effects of a dithiocarbamate fungicide, thiram (35 and 170 µg l(-1)), and of a petroleum middle distillate (0.01, 0.4, 2 and 20 mg l(-1)) in outdoor stream and pond mesocosms. Trait sensitivity was characterized using functional diversity indices and trait modality distributions to assess the influence of the type of experimental systems and the ability of traits to disentangle chemical-induced effects from temporal and stochastic variations. In addition, leaf litter breakdown was used as an integrative functional endpoint. Regardless to the substance, treatments had a direct effect on the functional structure of benthic macroinvertebrate communities in streams but not in ponds, suggesting that global functional responses to chemicals are system-specific. Although both substances had an effect in streams, differences were noticed in the nature of the affected traits suggesting that chemical mode of action plays a role in functional alterations. This was illustrated by the link between negative effects of chemical exposure on detritivorous taxa and reduced litter breakdown rate in streams. Therefore, characterisation of macroinvertebrate biological traits associated with the measurement of a functional process such as litter breakdown may provide a comprehensive understanding of the effects occurring in mesocosms exposed to organic chemicals.

Structural and biological trait responses of diatom assemblages to organic chemicals in outdoor flow-through mesocosms.

The sensitivity of diatom taxonomy and trait-based endpoints to chemicals has been poorly used so far in Environmental Risk Assessment. In this study, diatom assemblages in outdoor flow-through mesocosms were exposed to thiram (35 and 170 µg/L), and a hydrocarbon emulsion (HE; 0.01, 0.4, 2 and 20 mg/L). The effects of exposure were assessed for 12 weeks, including 9 weeks post-treatment, using taxonomic structure and diversity, bioindication indices, biological traits, functional diversity indices, indicator classes and ecological guilds. For both chemicals, diversity increased after the treatment period, and responses of ecological traits were roughly identical with an abundance increase of motile taxa tolerant to organic pollution and decrease of low profile taxa. Bioindication indices were not affected. Traits provided a complementary approach to biomass measurements and taxonomic descriptors, leading to a more comprehensive overview of ecological changes due to organic chemicals, including short- and long-term effects on biofilm structure and functioning.

Isotopic niche metrics as indicators of toxic stress in two freshwater snails.

Descriptors of trophic niche and of food web structure and function have been suggested as integrative and sensitive endpoints of toxicant effects. In the present study, carbon and nitrogen stable isotope signatures were used to assess the effects of the dithiocarbamate fungicide thiram (35 and 170µg/L nominal concentrations) and of a petroleum distillate (0.01, 0.4, 2 and 20mg/L nominal loadings as Hydrocarbon Emulsion or Hydrocarbon Water Accommodated Fraction) on the trophic niche of two freshwater gastropods in artificial streams (Radix peregra) and ponds (Lymnaea stagnalis). Results were analyzed using classical univariate statistical methods and recently proposed uni- and multivariate metrics of the realized trophic niche of species. The trophic niche metrics were highly sensitive to both types of chemicals, but exposure resulted in different response patterns according to the nature of the tested compound. Thiram clearly affected gastropod trophic niche leading to a change in the food resources used and resulting in trophic niche expansion (i.e., increase of diversity of used resources, especially dead animals) or trophic niche contraction (i.e., decrease of diversity of used resources) across time. Both gastropod taxa exposed to hydrocarbons showed a clear trophic niche expansion. Trophic niche metrics therefore provide a promising way of investigating non-lethal effects of exposure to organic chemicals on aquatic invertebrates, and subsequent disturbances in food webs.

Secondary production of freshwater zooplankton communities exposed to a fungicide and to a petroleum distillate in outdoor pond mesocosms.

Ecological risk assessment of chemicals in mesocosms requires measurement of a large number of parameters at the community level. Studies on invertebrate communities usually focus on taxonomic approaches, which only provide insights into taxonomic structure changes induced by chemicals. In the present study, abundance, biomass (B), theoretical production (P), and instantaneous P/B ratio were used as endpoints to assess the effects of the commercial form of the dithiocarbamate fungicide thiram (35 µg/L and 170 µg/L nominal concentrations) and of the hydrocarbon water accommodated fraction (HWAF) of a petroleum distillate (0.01 mg/L, 0.4 mg/L, 2 mg/L, and 20 mg/L loadings) on the zooplankton community in freshwater pond mesocosms. Endpoints were measured during a 4-wk treatment period (1 pulse/wk) followed by a 5-mo posttreatment period to evaluate zooplankton population recovery. The chlorophyll a concentration in water was significantly increased after treatment with HWAF, whereas it was not affected by thiram treatment. Zooplankton abundance-based analysis showed effects on a limited number of taxa, whereas other endpoints (mainly the P/B ratio) revealed that more taxa were impacted, with recovery depending on the chemical and concentration. Exposure to HWAF mainly had a negative impact on cladocerans, which resulted in top-down effects (between cladocerans and phytoplankton). Thiram negatively affected rotifers and copepods, suggesting more direct toxic effects. The results show that the use of secondary production as an endpoint provides a more comprehensive assessment of potential direct and indirect effects of chemicals on a community, and they also support evidence of alteration in functional processes.

Changes in the swimming behavior of Eurytemora affinis (Copepoda, Calanoida) in response to a sub-lethal exposure to nonylphenols.

Estuarine waters are continuously loaded with chemicals which affect the physiology of aquatic organisms to various extents. They also have adverse effects on a wide range of behaviors. Nonylphenols and related compounds are biodegradation products of the nonionic surfactants nonylphenol polyethoxylates. They are commonly found in the aquatic environment. We observed immediate alterations of the free swimming activity of the calanoid copepod Eurytemora affinis from the Seine estuary in response to a sub-lethal concentration of nonylphenols (4-NP and NP1EC) at environmentally realistic concentrations (2 µg/L). Swimming speed and activity increased for both males and females. The use of copepod kinematic proved to be a sensitive indicator of sub-lethal exposure to pollutants.

Uptake and elimination, and effect of estrogen-like contaminants in estuarine copepods: an experimental study.

BACKGROUND, AIM, AND SCOPE: In recent years, anthropogenic chemicals which can disrupt the hormonal systems of both humans and wildlife have been raised to a major cause of concern. The aim of the present work was to determine the bioconcentration factors of the two major alkylphenols (AP) of the Seine Estuary [4-nonylphenol (4 NP) and nonylphenol acetic acid (NP1EC)] and of the synthetic estrogen, estrogen ethinylestradiol (EE2), in Eurytemora affinis after exposure in a continuous flow-through system under environmental realistic conditions. Moreover, the elimination of these compounds in copepods from the Seine Estuary has been investigated by measuring concentrations after 1 week in clean water in comparison to background levels. MATERIALS AND METHODS: In this study, the dominant copepod species of the Seine Estuary, E. affinis, was exposed at environmental relevant concentrations under laboratory-controlled conditions to selected waterborn contaminants, a mixture of 4 NP/NP1EC, and a synthetic EE2. The uptake and the elimination of these contaminants by E. affinis have been studied. RESULTS: The results show that, at the end of the uptake period, both 4 NP and NP1EC, and also EE2 were accumulated in exposed copepods with respective concentration factors of 324, 3,020, and 5,383. A rapid elimination of these compounds was also observed in copepods placed in clean water since 54% of total NP1EC and 100% of EE2 amounts have been lost after 3 days. Pregnenolone was synthesized after exposure to EE2 and AP mixture. DISCUSSION: These results demonstrate that E. affinis has the potency to accumulate but also to eliminate endocrine disrupters which suggests a non-negligible role of this copepod species in the biogeochemical cycles of these contaminants in estuarine ecosystems. Hence, these results also suggest that a transfer of 4 NP, NP1EC, and EE2 to copepod predators and subsequently that secondary poisoning of these organisms might be possible. Estrogen-like contaminants can induce pregnenolone synthesis and affect the reproduction of E. affinis. CONCLUSIONS: These results suggest the important role of this copepod species in biogeochemical cycles of non-ionic surfactants as well as synthetic steroids in estuarine ecosystems. RECOMMENDATIONS AND PERSPECTIVES: E. affinis could be a non-negligible route of exposure for juvenile fish and underline the potential for deleterious effects on copepod predators.

Uptake and elimination of hydrophobic organic contaminants in estuarine copepods: an experimental study.

Polycyclic aromatic hydrocarbons (PAHs) are considered to be rapidly biotransformed by organisms, whereas polychlorinated biphenyls (PCBs) are strongly bioaccumulated. In the present study, the estuarine copepod Eurytemora affinis was exposed in a continuous flow-through system to dissolved PAH (500 ng/L) and PCB (300 ng/L) mixtures for 86 h, whereas control groups were placed in a continuous flow-through system with clean water. Both PCB and PAH body residues were measured and compared in exposed and in nonexposed copepods to assess the uptake and the elimination of these two contaminant classes in this copepod species. After the exposure, exposed copepods exhibited concentration factors, based on a dry-weight basis, of 25, 750, and 1,200, respectively, for total PCBs and PAHs. The lower concentrations of PAHs in the nonexposed versus exposed copepods in contrast to small differences for PCBs suggest a higher rate of metabolism of PAHs compared with PCBs and could explain the differences observed in the accumulation. Furthermore, uptake as well as elimination of both PCBs and PAHs were compound selective in E. affinis. Therefore, higher-molecular-weight PCBs and PAHs were preferentially accumulated, while lower-molecular-weight compounds were preferentially eliminated. These results suggest the importance of copepods in the biogeochemical cycles of hydrophobic organic contaminants in estuarine ecosystems.