Frédéric Orias

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.

Tamoxifen ecotoxicity and resulting risks for aquatic ecosystems

Tamoxifen, a drug used to treat cancer, is regularly found in hydrosystems at concentrations of several hundred ng L(-1). To characterize its ecotoxicity, we implemented a battery of bioassays on organisms belonging to 3 different trophic levels: Pseudokirchneriella subcapitata, Chlorella vulgaris and Chlamydomonas reinhardtii, for primary producers, Daphnia magna (immobilization, grazing and reproduction) for primary consumers, and Danio rerio for secondary consumers (embryotoxicity test). In view of the results obtained and the ecotoxicity values of tamoxifen available in the literature, we established a PNEC (Predictive No Effect Concentration) equal to 81 ng L(-1) for continental water. This PNEC allowed us to calculate Risk Quotients (RQ) for 4 continental hydrosystems in 4 different countries in which measures of tamoxifen had already been performed on surface waters. In two of the situations studied, RQs were higher than 1, reaching a maximum of 2.6. These results show the need to deepen the characterization of ecotoxicological risks linked to the discharge of tamoxifen in surface waters. In addition, we propose applying this approach to other drug residues detected in the environment.

Experimental assessment of the bioconcentration of 15N-tamoxifen in Pseudokirchneriella subcapitata

Nowadays, pharmaceutical compounds (PC) are ubiquitous in aquatic ecosystems. In addition to direct ecotoxicity, the bioconcentration of PC in organisms is a phenomenon which could have an impact on the whole ecosystem. In order to study this phenomenon, we exposed unicellular algae (Pseudokirchneriella subcapitata) to (15)N-tamoxifen, an anticancer drug labelled with a stable nitrogen isotope used as a tracer. By measuring (15)N enrichment over time, we were able to measure the increase of tamoxifen content in algae. This enrichment was measured by an elemental analyser coupled with an isotopic ratio mass spectrometer (EA-IRMS). Algal cells were exposed for 7d to 3 concentrations of tamoxifen: 1, 10 and 100μgL(-1). Our result shows a high bioconcentration in algae from the first minutes of contact. The highest bioconcentration factor measured is around 26500. We also observe that bioconcentration is not linked to the exposure concentration. This study is the first to use stable isotopes in order to monitor PCs in aquatic organisms such as algae. The use of stable isotopes in ecotoxicology offers interesting perspectives in the field of contaminant transfer in organisms and along the trophic web.

Bioconcentration of 15N-tamoxifen at environmental concentration in liver, gonad and muscle of Danio rerio

Pharmaceutical compounds (PCs) are ubiquitous in aquatic ecosystems. In addition to the direct ecotoxicological risk presented by certain PCs, others can accumulate inside organisms and along trophic webs, subsequently contaminating whole ecosystems. We studied the bioconcentration of a bioaccumulative PC already found several times in the environment: tamoxifen. To this end, we exposed Danio rerio for 21d to (15)N-tamoxifen concentrations ranging from 0.1 to 10µg/L and used an analytic method based on stable isotopes to evaluate the tamoxifen content in these organisms. The evolution of the (15)N/(14)N ratio was thus measured in liver, muscle and gonads of exposed fish compared to control fish. We succeeded in quantifying (15)N-tamoxifen bioconcentrations at all the exposure concentrations tested. The highest bioconcentration factors of tamoxifen measured were 14,920 in muscle, 73,800 in liver and 85,600 in gonads of fish after 21d exposure at a nominal concentration of 10µg/L. However, these bioconcentration factors have to be considered as maximal values (BCFMAX). Indeed, despite its proven stability, tamoxifen can be potentially partially degraded during experiments. We now need to refine these results by using a direct analytic method (i.e. LC-MS/MS).

Co-encapsulation of Daphnia magna and microalgae in silica matrices, a stepping stone toward a portable microcosm

We report on the first silica encapsulation of a metazoan (Daphnia magna), with a high initial viability (96% of the population remained active 48 h after encapsulation). Moreover, the co-encapsulation of this crustacean and microalgae (Pseudokirchneriella subcapitata) was achieved, creating inside a silica monolith, the smallest microcosm developed to present. This artificial ecosystem in a greatly diminished scale isolated inside a silica nanoporous matrix could have applications in environmental monitoring, allowing ecotoxicity studies to be carried out in portable devices for on-line and in situ pollution level assessment.

Ecotoxicity of Hospital Wastewater

During the last 10 years, characterizing the ecotoxicity of hospital wastewater (HWW) has focused on different aspects. Initially, it mainly consisted in collecting information on ecotoxic substances used in hospitals (disinfectants, detergents, pharmaceuticals, etc.). Thereafter, experimental measurements of ecotoxicity on whole effluent were carried out in different hospitals. These data have shown the generally high ecotoxicity of this effluent, and its considerable evolution during a single day and a full year of activity. In addition, the bioaccumulation of certain pharmaceuticals in organisms and trophic chains has been demonstrated, which contributes to increasing the risk of these molecules. The interactions between the molecules present in HWW have also been the subject of studies. The collection of all these data have enabled researchers and managers to carry out the first ecotoxicological risk assessment studies in different hospitals around the world. It is now necessary to complete these works, in order to refine the characterization of HWW ecotoxicity and consolidate the methodologies of ecotoxicological risk assessment formulated.