SIPIBEL observatory: Data on usual pollutants (solids, organic matter, nutrients, ions) and micropollutants (pharmaceuticals, surfactants, metals), biological and ecotoxicity indicators in hospital and urban wastewater, in treated effluent and sludge from wastewater treatment plant, and in surface and groundwater.

The Bellecombe pilot site - SIPIBEL - was created in 2010 in order to study the characterisation, treatability and impacts of hospital effluents in an urban wastewater treatment plant. This pilot site is composed of: i) the Alpes Léman hospital (CHAL), opened in February 2012, ii) the Bellecombe wastewater treatment plant, with two separate treatment lines allowing to fully separate the hospital wastewater and the urban wastewater, and iii) the Arve River as the receiving water body and a tributary of the Rhône River and the Geneva aquifer. The database includes in total 48 439 values measured on 961 samples (raw and treated hospital and urban wastewater, activated sludge in aeration tanks, dried sludge after dewatering, river and groundwater, and a few additional campaigns in aerobic and anaerobic sewers) with 44 455 physico-chemistry values (including 15 pharmaceuticals and 14 related transformation products, biocides compounds, metals, organic micropollutants), 2 193 bioassay values (ecotoxicity), 1 679 microbiology values (including microorganisms and antibioresistance indicators) and 112 hydrobiology values.

Ecotoxicity and antibiotic resistance of wastewater during transport in an urban sewage network.

Urban wastewater (UWW) management usually entails biological and physicochemical monitoring due to its potential impact on the quality of the receiving environment. A major component of a sewage system is the pipe network leading the water to the treatment plant. Up to now, few studies have been conducted on the diverse phenomena that may affect the characteristics of the water during its transportation. In this study, ecotoxicity and potential antibiotic resistance were used in a global method to assess the change of UWW quality in a sewage system and determine if sewer pipes can act as a bioreactor spread. Three bioassays were conducted to assess the ecotoxicity of the samples and the concentration and relative abundance of two classes of integrons (as a proxy for antibiotic resistance) were measured. The results of the bioassay battery do not show a pattern, despite the fact that differences were noticeable between upstream and downstream samples. Antibiotic resistance appeared to decrease during transport in the pipe as the concentration and relative abundance of integrons decreased during several campaigns. This result should be confirmed in other sewer networks but already provides useful information for the management of urban sewage system.

Microalgal whole-cell biomarkers as sensitive tools for fast toxicity and pollution monitoring of urban wet weather discharges.

Urban wet weather discharge (UWWD) management is an important issue. UWWD often represents a significant source of pollution in all aquatic bodies. The occurrence of this pollution is difficult to predict due to the variability of storm events and the unknown contents of urban watershed leached out by rain. Previous studies have tried to demonstrate the ecotoxic impact of UWWD. However, most of them merely highlight the limitations of classic monospecific bioassays, given the high dilution of micropollutants or the presence of nutrients masking toxic effects. Overcoming this problem is therefore of great interest. In this study, we demonstrated the utility of a battery of biomarkers (e.g. membrane permeability, chlorophyll fluorescence, esterase and alkaline phosphatase activities) on the microalgae Chlorella vulgaris to detect the toxic effects of 7 UWWD samples after short exposures (2 and 24 h). These biomarkers are linked to microalgal life traits. Complementarily, monospecific bioassays were carried on Pseudokirchneriella subcapitata, Chlorella vulgaris, Daphnia magna and Heterocypris incongruens to compare their sensitivity to the UWWD samples. No toxic effect was detected in any of the bioassays. Yet, algal biomarkers indicated a disturbance in microalgae physiology, and particularly a perturbation of chlorophyll fluorescence, which was observed in all of the samples tested. While algal membrane permeability was affected by only one UWWD, these two enzymatic activities were stimulated or inhibited depending on the sample. Finally, this study demonstrates the sensitivity of algal biomarkers and the need to develop new, fruitful approaches to characterizing UWWD toxicity.

Ecotoxicity and antibiotic resistance of a mixture of hospital and urban sewage in a wastewater treatment plant.

Hospital and urban effluents are a source of diverse pollutants such as organic compounds, heavy metals, detergents, disinfectants, pharmaceuticals, and microorganisms resistant to antibiotics. Usually, these two types of effluent are mixed in the sewage network, but a pilot site in France now allows studying them separately or mixed to understand more about their characteristics and the phenomena that occur following their mixing. In this study, their ecotoxicity (Daphnia magna mobility, Pseudokirchneriella subcapitata growth, Brachionus calyciflorus reproduction, and SOS Chromotest) and antibiotic resistance (integron quantification) were assessed during mixing and treatment steps. The main results of this study are (i) the ecotoxicity and antibiotic resistance potentials of hospital wastewater are higher than in urban wastewater and (ii) mixing two different effluents does not lead to global synergistic or antagonistic effects on ecotoxicity and antibiotic resistance potential. The global additivity effect observed in this case must be confirmed by other studies on hospital and urban effluents on other sites to improve knowledge relating to this source of pollution and its management.

Spatial and temporal dynamics of sediment ecotoxicity in urban stormwater retention basins: Methodological approach and application to a pilot site close to Lyon in France.

To characterize the spatio-temporal variation of sediment ecotoxicity in a retention/detention basin, a monitoring program using the Heterocypris incongruens bioassay was carried out for 72 months (5 years) on a field basin close to Lyon in France. Results showed that the variation of ecotoxicity is relatively small from one location of the basin to another, apart from sediment sampling collected in an open-air chamber located in basin supposed to collect gross pollutants and hydrocarbons. Regarding the temporal variation of ecotoxicity, the bioassays also showed a slight variation between 6 and 72 months. On the contrary, they highlighted the high ecotoxicity of the "fresh" sediments collected during rain events using sediment traps. Additional investigations are needed to understand the period of inflexion of ecotoxicity, occurring between 24 h and 6 months. These results can be used by practitioners of urban facilities and networks to improve maintenance strategies of retention/detention basins.

Assessment of long term ecotoxicity of urban stormwaters using a multigenerational bioassay on Ceriodaphnia dubia: A preliminary study.

Standardized ecotoxicity bioassays show some limits to assess properly long-term residual toxicity of complex mixture of pollutants often present at low concentration, such as stormwaters. Among invertebrate organisms used for ecotoxicity testing, the microcrustacean Ceriodaphnia dubia (C. dubia) is considered as one of the most sensitive, especially regarding reproduction impairment as a toxicity endpoint. Consequently, this work explores the interest to perform a multigenerational assay based on the study of the reproduction of C. dubia to assess long-term ecotoxicity of complex mixture, using stormwater samples. With this in mind, a battery of standardized bioassays (Daphnia magna mobility, Pseudokirchneriella subcapitata population growth, Heterocypris incongruens growth and one generation C. dubia reproduction inhibition assays) was performed in parallel to a three generation C. dubia reproduction inhibition assay on 2 stormwater samples. Results highlighted that while all standardized bioassays failed to reveal residual toxicity in the stormwater samples, the C. dubia multigenerational assay exhibited an higher sensitivity than the previous ones. No adverse effect was observed for the first exposed generation, but an increase in mortality and a reproduction disturbance was obtained in the second and third exposed generation depending of the sample. Further experiments are now needed to optimize the exposure protocol of this multigenerational assay.

A posteriori assessment of ecotoxicological risks linked to building a hospital.

Hospital wastewater (HWW) contain a large number of chemical pollutants such as disinfectants, surfactants, and pharmaceutical residues. A part of these pollutants is not eliminated by traditional urban wastewater treatment plants (WWTP), leading to a risk for the aquatic ecosystems receiving these effluents. In order to assess this risk, we formulated a specific methodology based on the ecotoxicological characterisation of the hospital wastewater using a battery of three chronic bioassays (Pseudokirchneriella subcapitata, Heterocypris incongruens and Brachionus calyciflorus). We used it for the posteriori risk assessment of a hospital recently built in south-east France, and we studied the evolution of this risk during two years. We also used it to assess the decrease of the ecotoxicological risk after treatment of the effluent in a specific line of the local WWTP. Lastly, we compared these results with the risk assessment made before the building of the hospital in the context of a priori risk assessment. The results obtained showed an important evolution of the risk overtime, according to the hospital activities and the river flows, and a real decrease of the risk after treatment in the dedicated line. They also showed that the a priori assessment of ecotoxicological risks, made previously, was overstated, mainly because of the application of the precautionary principle.

Proposal to optimize ecotoxicological evaluation of wastewater treated by conventional biological and ozonation processes.

A mixture of urban and hospital effluents (50% v/v) was evaluated for ecotoxicity with an advanced bioassay battery. Mixed effluents were tested before any treatment, after biological treatment alone, and after biological treatment followed by a tertiary ozonation (15 mg O3/L). Laying a high value on the continuance of organisms' fitness, essential to preserve a healthy receiving ecosystem, the main objective of this study was to combine normalized bioassays with newly developed in vivo and in vitro tests in order to assess alteration of embryo development, growth and reproduction, as well as genotoxic effects in aquatic organisms exposed to complex wastewater effluents. Comparison of the bioassays sensitivity was considered. Contrary to the lack of toxicity observed with normalized ecotoxicity tests, endpoints measured on zebrafish embryos such as developmental abnormalities and genotoxicity demonstrated a residual toxicity in wastewater both after a biological treatment followed or not by a tertiary O3 treatment. However, the ozonation step allowed to alleviate the residual endocrine disrupting potential measure in the biologically treated effluent. This study shows that normalized bioassays are not sensitive enough for the ecotoxicological evaluation of wastewaters and that there is a great need for the development of suitable sensitive bioassays in order to characterize properly the possible residual toxicity of treated effluents.

Spatial variability of sediment ecotoxicity in a large storm water detention basin.

Detention basins are valuable facilities for urban storm water management, from both the standpoint of flood control and the trapping of pollutants. Studies performed on storm water have shown that suspended solids often constitute the main vector of pollutants (heavy metals, polycyclic aromatic hydrocarbon (PAH), etc.). In order to characterise the ecotoxicity of urban sediments from storm water detention basins, the sediments accumulated over a 6-year period were sampled at five different points through the surface of a large detention basin localised in the east of Lyon, France. A specific ecotoxicological test battery was implemented on the solid phase (raw sediment) and the liquid phase (interstitial water of sediments). The results of the study validated the method formulated for the ecotoxicological characterization of urban sediments. They show that the ecotoxicological effect of the sediments over the basin is heterogeneous and greater in areas often flooded. They also show the relationship between, on one hand, the physical and chemical characteristics of the sediments and, on the other hand, their ecotoxicity. Lastly, they contribute to a better understanding of the dynamics of the pollution close to the bottom of detention basins, which can be useful for improving their design. The results of this research raise particularly the issue of using oil separators on the surface of detention basins.

Inter-laboratory trial of a standardized sediment contact test with the aquatic plant Myriophyllum aquaticum (ISO 16191).

A whole-sediment toxicity test with Myriophyllum aquaticum has been developed by the German Federal Institute of Hydrology and standardized within the International Organization for Standardization (ISO; ISO 16191). An international ring-test was performed to evaluate the precision of the test method. Four sediments (artificial, natural) were tested. Test duration was 10 d, and test endpoint was inhibition of growth rate (r) based on fresh weight data. Eighteen of 21 laboratories met the validity criterion of r ≥ 0.09 d(-1) in the control. Results from 4 tests that did not conform to test-performance criteria were excluded from statistical evaluation. The inter-laboratory variability of growth rates (20.6%-25.0%) and inhibition (26.6%-39.9%) was comparable with the variability of other standardized bioassays. The mean test-internal variability of the controls was low (7% [control], 9.7% [solvent control]), yielding a high discriminatory power of the given test design (median minimum detectable differences [MDD] 13% to 15%). To ensure these MDDs, an additional validity criterion of CV ≤ 15% of the growth rate in the controls was recommended. As a positive control, 90 mg 3,5-dichlorophenol/kg sediment dry mass was tested. The range of the expected growth inhibition was proposed to be 35 ± 15%. The ring test results demonstrated the reliability of the ISO 16191 toxicity test and its suitability as a tool to assess the toxicity of sediment and dredged material.

Evaluation of ecotoxicological risks related to the Discharge of Combined Sewer Overflows (CSOs) in a periurban River.

Discharges of Combined Sewer Overflows (CSOs) into periurban rivers present risks for the concerned aquatic ecosystems. In this work, a specific ecotoxicological risk assessment methodology has been developed as management tool to municipalities equipped with CSOs. This methodology comprises a detailed description of the spatio-temporal system involved, the choice of ecological targets to be preserved, and carrying out bioassays adapted to each compartment of the river receiving CSOs. Once formulated, this methodology was applied to a river flowing through the outskirts of the city of Lyon in France. The results obtained for the scenario studied showed a moderate risk for organisms of the water column and a major risk for organisms of the benthic and hyporheic zones of the river. The methodology enabled identifying the critical points of the spatio-temporal systems studied, and then making proposals for improving the management of CSOs.

Assessment of ecological risks linked to the discharge of saline industrial effluent into a river.

Discharges of saline effluents into rivers can lead to risks for local aquatic ecosystems. A specific ecological risk assessment methodology has been developed to propose a management tool to organisations responsible for managing rivers and industrial companies producing saline effluents. This methodology involves the detailed description of the spatiotemporal system concerned, the choice of ecological targets to be preserved, and the performance of bioassays adapted to each of the compartments of the river. Following development, it was applied to an industrial effluent in eastern France. For the scenario studied, results obtained suggest a high risk for the organisms of the water column and a low risk for the organisms of the periphyton. This difference can be explained by the structure of the latter which integrate extracellular polymers secreted by the organisms of the biofilm, forming a gel with a porous structure that acts as a barrier to diffusion. The methodology formulated permitted identifying the critical points of the spatiotemporal system studied and then using them as the basis for making well-grounded proposals for management. Lastly, proposals to improve the methodology itself are made, especially concerning the integration of the sediment compartment in the version formulated initially.

Ecotoxicological risk assessment linked to infilling quarries with treated dredged seaport sediments.

The dredged sediments of polluted seaports now raise complex management problems since it is no longer possible to discharge them into the sea. This results in the need to manage them on land, raising other types of technical, economic and environmental problems. Regarding the technical and economic dimensions, traditional waste treatment methods have proved to be poorly adapted, due to very high costs and low absorbable volumes. In this context, filling quarries in coastal areas with treated sediments could represent an interesting alternative for these materials. Nevertheless, for the environmental dimension, it is necessary to demonstrate that this possibility is harmless to inland ecosystems. Consequently, a specific ecotoxicological risk assessment methodology has been formulated and tested on three sediments taken from seaboards of France, in view to providing an operational and usable tool for the prior validation of any operation to fill quarries with treated seaport sediments. This method incorporates the formulation of a global conceptual model of the scenario studied and the definition of protocols for each of its steps: the characterisation of exposures (based on a simulation of sediment deposit), the characterisation of effects (via the study of sediments ecotoxicity), and the final ecotoxicological risk assessment performed as a calculation of a risk quotient. It includes the implementation in parallel of two types of complementary approach: the "substances" approach derived from the European methodology for assessing new substances placed on the market, and the "matrix" approach which is similar to methods developed in France to assess ecological risks in other domains (waste management, polluted site management, …). The application of this dual approach to the three sediments tested led to conclude with reliability that the project to deposit sediments "1" and "2" presented a low risk for the peripheral aquatic ecosystems while sediment "3" presented a high risk.

Fate of glutaraldehyde in hospital wastewater and combined effects of glutaraldehyde and surfactants on aquatic organisms.

Glutaraldehyde (GA), an aliphatic dialdehyde disinfectant, and surfactants, one of the major components of detergents, are widely used in hospitals in order to eliminate pathogenic organisms causing nosocomial infectious diseases. After their use, disinfectants and surfactants reach the wastewater network together. The discharge of chemical compounds from hospital activities into wastewater is also a well-known problem, causing pollution of water resources and constituting an ecological risk for aquatic organisms. In this study, the chemistry and toxicology of GA and surfactant mixtures were reviewed in order to estimate their fate in aquatic ecosystems. Furthermore, their joint effects on aquatic organisms were experimentally assessed in the laboratory. A simple model of the additive joint action of toxicants was used to determine combined acute toxicity effects on the bacteria luminescence and Daphnia mobility of three mixtures containing GA at 1.5 x EC50 24 h [in mg/L] on Daphnia and anionic, cationic and nonionic surfactants at twice their critical micellar concentration (CMC). The mixture of GA and a cationic surfactant gave an EC50 30 min on Vibrio fischeri of 0.158%, with a concentration of 0.04 mg GA/L and 1.04 mg CTAB/L, which provided an additive action. The interaction between GA and an anionic surfactant on V. fischeri produced an antagonistic joint action with an EC50 30 min of 3.95%, containing 1.06 mg GA/L and 33.2 mg SDS/L. A synergistic action with an EC50 30 min of 8.4% on V. fischeri was observed for the mixture containing GA and a nonionic surfactant. Antagonistic interactions were observed for the joint action between GA and the surfactants studied on Daphnia. The mixture of GA and CTAB was more toxic (EC50 24 h=0.02%) than the two other mixtures (EC50 24 h GA+SDS=6%; EC50 24 h GA+TX 100=10%). This study provides new data on the toxicity of certain hospital pollutants entering the aquatic environment and detected in surface and groundwaters. It is necessary to study the joint effects of GA and surfactant mixtures following chronic and sublethal standard bioassays in order to estimate the contribution of the additive joint action models in assessing the environmental risk of hospital wastewater (HW).