International audience | Characterizing waste ecotoxicity is laborious because of both the undefined nature of environmental samples and the diversity of contaminants that can be present. With regard to these limitations, traditional approaches do not provide information about the nature of the pollution encountered. To improve such assessments, a fluorescent library of 1870 transcriptomic reporters from Escherichia coli K12 MG1655 was used to report the ecotoxic status of environmental samples. The reliability of the approach was evaluated with 6 metallic pollutants (As, Cu, Cd, Hg, Pb, Zn) used alone and in mixture in pure and complex matrices. A total of 18 synthetic samples were used to characterize the specificity of the resulting metallic contamination fingerprints. Metallic contamination impacted 4.5 to 10.2% of the whole transcriptomic fingerprint of E. coli. The analysis revealed that a subset of 175 transcriptomic reporters is sufficient to characterize metallic contamination, regardless of the nature of the sample. A statistical model distinguished patterns due to metallic contamination and provided information about the level of toxicity with 93 to 98% confidence. The use of the transcriptomic assessment was validated for 17 complex matrices with various toxicities and metal contaminants, such as activated sludge, wastewater effluent, soil, wood and river water. The presence of metals and their associated toxicity, which seems linked to their bioavailabilities, were thereby determined. This method constitutes a possible tool to screen unknown complex samples for their metallic status and identify those for which a deeper characterization must be achieved by the use of traditional biosensors and analytical methods.

Characterizing waste ecotoxicity is laborious because of both the undefined nature of environmental samples and the diversity of contaminants that can be present. With regard to these limitations, traditional approaches do not provide information about the nature of the pollution encountered. To improve such assessments, a fluorescent library of 1870 transcriptomic reporters from Escherichia coli K12 MG1655 was used to report the ecotoxic status of environmental samples. The reliability of the approach was evaluated with 6 metallic pollutants (As, Cu, Cd, Hg, Pb, Zn) used alone and in mixture in pure and complex matrices. A total of 18 synthetic samples were used to characterize the specificity of the resulting metallic contamination fingerprints. Metallic contamination impacted 4.5 to 10.2% of the whole transcriptomic fingerprint of E. coli. The analysis revealed that a subset of 175 transcriptomic reporters is sufficient to characterize metallic contamination, regardless of the nature of the sample. A statistical model distinguished patterns due to metallic contamination and provided information about the level of toxicity with 93 to 98% confidence. The use of the transcriptomic assessment was validated for 17 complex matrices with various toxicities and metal contaminants, such as activated sludge, wastewater effluent, soil, wood and river water. The presence of metals and their associated toxicity, which seems linked to their bioavailabilities, were thereby determined. This method constitutes a possible tool to screen unknown complex samples for their metallic status and identify those for which a deeper characterization must be achieved by the use of traditional biosensors and analytical methods.

International audience | Some steps of the soil nitrogen (N) cycle are sensitive to environmental pressures like soil moisture or contamination, which are expected to evolve during the next decades. Individual stresses have been well studied, but their combination is not yet documented. In this work, we aimed at assessing the importance of the soil moisture on the impact of copper (Cu) contaminations on the N cycling soil function using the potential nitrification activities (PNA) as bioindicator. A two-step experiment was performed. First, a loamy soil was incubated 5 weeks in either 30, 60, or 90% of its water holding capacity (WHC) or alternating drought and rewetting periods. Thereafter, soil samples were exposed to a gradient of Cu concentrations through a bioassay involving nitrification. The dose-response curves of PNA in function of added Cu were modeled to calculate the effective Cu concentrations, namely ECx with x being the percentage of PNA inhibition. These values were then compared between experimental conditions to highlight differences in threshold values. The preincubation moisture treatments significantly affected the PNA responses to the secondary Cu stress with, for instance, hormetic responses in all cases except for the dry-rewetting treatment. Small PNA inhibitions were estimated for high Cu doses in the soils with low water contents (30% WHC) or submitted to dry-rewetting cycles, contrarily to the patterns observed for the soils with high water contents (90% WHC) or submitted to a single period of drought. Overall, significant differences were found in estimated ECx values between moisture treatments.

International audience | Some steps of the soil nitrogen (N) cycle are sensitive to environmental pressures like soil moisture or contamination, which are expected to evolve during the next decades. Individual stresses have been well studied, but their combination is not yet documented. In this work, we aimed at assessing the importance of the soil moisture on the impact of copper (Cu) contaminations on the N cycling soil function using the potential nitrification activities (PNA) as bioindicator. A two-step experiment was performed. First, a loamy soil was incubated 5 weeks in either 30, 60, or 90% of its water holding capacity (WHC) or alternating drought and rewetting periods. Thereafter, soil samples were exposed to a gradient of Cu concentrations through a bioassay involving nitrification. The dose-response curves of PNA in function of added Cu were modeled to calculate the effective Cu concentrations, namely ECx with x being the percentage of PNA inhibition. These values were then compared between experimental conditions to highlight differences in threshold values. The preincubation moisture treatments significantly affected the PNA responses to the secondary Cu stress with, for instance, hormetic responses in all cases except for the dry-rewetting treatment. Small PNA inhibitions were estimated for high Cu doses in the soils with low water contents (30% WHC) or submitted to dry-rewetting cycles, contrarily to the patterns observed for the soils with high water contents (90% WHC) or submitted to a single period of drought. Overall, significant differences were found in estimated ECx values between moisture treatments.

Some steps of the soil nitrogen (N) cycle are sensitive to environmental pressures like soil moisture or contamination, which are expected to evolve during the next decades. Individual stresses have been well studied, but their combination is not yet documented. In this work, we aimed at assessing the importance of the soil moisture on the impact of copper (Cu) contaminations on the N cycling soil function using the potential nitrification activities (PNA) as bioindicator. A two-step experiment was performed. First, a loamy soil was incubated 5 weeks in either 30, 60, or 90% of its water holding capacity (WHC) or alternating drought and rewetting periods. Thereafter, soil samples were exposed to a gradient of Cu concentrations through a bioassay involving nitrification. The dose–response curves of PNA in function of added Cu were modeled to calculate the effective Cu concentrations, namely ECx with x being the percentage of PNA inhibition. These values were then compared between experimental conditions to highlight differences in threshold values. The preincubation moisture treatments significantly affected the PNA responses to the secondary Cu stress with, for instance, hormetic responses in all cases except for the dry-rewetting treatment. Small PNA inhibitions were estimated for high Cu doses in the soils with low water contents (30% WHC) or submitted to dry-rewetting cycles, contrarily to the patterns observed for the soils with high water contents (90% WHC) or submitted to a single period of drought. Overall, significant differences were found in estimated ECx values between moisture treatments.

International audience | Methotrexate is an antineoplastic folate analog of high environmental concern, due to its low biodegradability and toxicological properties. This study focused on its photodegradation under two irradiation conditions, aiming to be representative of environment (300-450 nm) and drinking water treatment (254 nm). The photodegradation experiments were conducted at two pH, to vary the methotrexate ionization state and to produce a large variety of transformation products (TPs). The degradation kinetics determined through LC-UV monitoring were contrasted according to pH and irradiation wavelength. However, the quantum yields were independent of ionization state at 254 nm and the changes in kinetics at higher wavelengths were attributed to a change in the degradation mechanism. The TPs formed during the reactions were identified by UHPLC-MS/MS, using both the positive and negative modes. Among the eleven proposed structures, five were described as methotrexate TPs for the first time. The TPs result from N-demethylation, glutamic acid oxidation, and C-N cleavage, all of them leading to further degraded photoproducts presenting modified or lost glutamic acid part. This was made possible thanks to the negative mode, which allowed the exploration of the glutamic acid moiety modifications. Cytotoxicity assessment on A549 cancer cells demonstrated that all photoproducts formed at pH 7 were less toxic than the parent compound.

Methotrexate is an antineoplastic folate analog of high environmental concern, due to its low biodegradability and toxicological properties. This study focused on its photodegradation under two irradiation conditions, aiming to be representative of environment (300–450 nm) and drinking water treatment (254 nm). The photodegradation experiments were conducted at two pH, to vary the methotrexate ionization state and to produce a large variety of transformation products (TPs). The degradation kinetics determined through LC-UV monitoring were contrasted according to pH and irradiation wavelength. However, the quantum yields were independent of ionization state at 254 nm and the changes in kinetics at higher wavelengths were attributed to a change in the degradation mechanism. The TPs formed during the reactions were identified by UHPLC-MS/MS, using both the positive and negative modes. Among the eleven proposed structures, five were described as methotrexate TPs for the first time. The TPs result from N-demethylation, glutamic acid oxidation, and C–N cleavage, all of them leading to further degraded photoproducts presenting modified or lost glutamic acid part. This was made possible thanks to the negative mode, which allowed the exploration of the glutamic acid moiety modifications. Cytotoxicity assessment on A549 cancer cells demonstrated that all photoproducts formed at pH 7 were less toxic than the parent compound.

International audience | The EcotoxicoMicYR group was initially composed of 4 Ph.D. students and 4 post-doctoral researchers. In brief, the EcotoxicoMicYR webinar took place three Monday afternoons in a row from November 22 to December 6, 2021. These three half-day webinars reached a success beyond our expectations with 25 countries and 41 presentations. Keynote lectures were delivered by Dr Fabio Roldan (Pontificia Universidad Javeriana, Colombia), Dr Belinda Ferrari (The University of New South Wales, Australia), and Dr Ahmed Tlili (Eawag, Switzerland). Their presentations provided an insight on latest research developments in the microbial ecotoxicology field and highlighted their specific contribution to this discipline. Twenty-two oral presentations and 16 pre-recorded presentations were diffused.

The EcotoxicoMicYR group was initially composed of 4 Ph.D. students and 4 post-doctoral researchers. In brief, the EcotoxicoMicYR webinar took place three Monday afternoons in a row from November 22 to December 6, 2021. These three half-day webinars reached a success beyond our expectations with 25 countries and 41 presentations. Keynote lectures were delivered by Dr Fabio Roldan (Pontificia Universidad Javeriana, Colombia), Dr Belinda Ferrari (The University of New South Wales, Australia), and Dr Ahmed Tlili (Eawag, Switzerland). Their presentations provided an insight on latest research developments in the microbial ecotoxicology field and highlighted their specific contribution to this discipline. Twenty-two oral presentations and 16 pre-recorded presentations were diffused.

The EcotoxicoMicYR group was initially composed of 4 Ph.D. students and 4 post-doctoral researchers. In brief, the EcotoxicoMicYR webinar took place three Monday afternoons in a row from November 22 to December 6, 2021. These three half-day webinars reached a success beyond our expectations with 25 countries and 41 presentations. Keynote lectures were delivered by Dr Fabio Roldan (Pontificia Universidad Javeriana, Colombia), Dr Belinda Ferrari (The University of New South Wales, Australia), and Dr Ahmed Tlili (Eawag, Switzerland). Their presentations provided an insight on latest research developments in the microbial ecotoxicology field and highlighted their specific contribution to this discipline. Twenty-two oral presentations and 16 pre-recorded presentations were diffused.

International audience | The presence of SARS-CoV-2 RNA has been extensively reported at the influent of wastewater treatment plants (WWTPs) worldwide, and its monitoring has been proposed as a potential surveillance tool to early alert of epidemic outbreaks. However, the fate of the SARS-CoV-2 RNA in the treatment process of WWTP has not been widely studied yet; therefore, in this study, we aimed to evaluate the efficiency of treatment processes in reducing SARS-CoV-2 RNA levels in wastewater. The treatment process of three WWTPs of the Parisian area in France was monitored on six different weeks over a period of 2 months (from April 14 to June 9, 2021). SARS-CoV-2 RNA copies were detected using digital polymerase chain reaction (dPCR). Investigation on the presence of variants of concern (Del69-70, E484K, and L452R) was also performed. Additionally, SARS-CoV-2 RNA loads in the WWTPs influents were expressed as the viral concentration in per population equivalent (PE) and showed a good correlation with French public health indicators (incidence rate). SARS-CoV-2 RNA loads were notably reduced along the water treatment lines of the three WWTPs studied (2.5-3.4 log reduction). Finally, very low SARS-CoV-2 RNA loads were detected in effluents (non-detected in over half of the samples) which indicated that the potential risk of the release of wastewater effluents to the environment is probably insignificant, in the case of WWTPs enabling an efficient biological removal of nitrogen.

The presence of SARS-CoV-2 RNA has been extensively reported at the influent of wastewater treatment plants (WWTPs) worldwide, and its monitoring has been proposed as a potential surveillance tool to early alert of epidemic outbreaks. However, the fate of the SARS-CoV-2 RNA in the treatment process of WWTP has not been widely studied yet; therefore, in this study, we aimed to evaluate the efficiency of treatment processes in reducing SARS-CoV-2 RNA levels in wastewater. The treatment process of three WWTPs of the Parisian area in France was monitored on six different weeks over a period of 2 months (from April 14 to June 9, 2021). SARS-CoV-2 RNA copies were detected using digital polymerase chain reaction (dPCR). Investigation on the presence of variants of concern (Del69-70, E484K, and L452R) was also performed. Additionally, SARS-CoV-2 RNA loads in the WWTPs influents were expressed as the viral concentration in per population equivalent (PE) and showed a good correlation with French public health indicators (incidence rate). SARS-CoV-2 RNA loads were notably reduced along the water treatment lines of the three WWTPs studied (2.5–3.4 log reduction). Finally, very low SARS-CoV-2 RNA loads were detected in effluents (non-detected in over half of the samples) which indicated that the potential risk of the release of wastewater effluents to the environment is probably insignificant, in the case of WWTPs enabling an efficient biological removal of nitrogen.

Although banned for years, organochlorine pesticides and PCBs continue to affect aquatic life, dolphins being particularly exposed. The concentrations of 31 PCB congeners, and 15 banned pesticides or metabolites were measured in 5 tissues of 68 striped dolphins stranded in the Northwestern Mediterranean coast in 2010–16. The results were compared to historical data (1988–2009) and, even though there is a slow decreasing trend, the levels in the 2010–2016 samples were still elevated based on common cetacean toxicological thresholds. A transition period in 2007–08, probably caused by a morbillivirus epizootic amplified the stranding, espacially of highly contaminated specimens. From 2010, higher proportions in parent compounds towards metabolites were observed yet again. These changing patterns were likely reflect the exposure of dolphins to the remobilization of pollutants from contaminated soils and sediments, with a prominent role of rivers. This should lead to an even slower decline of these contaminants that could last for decades, requiring new efforts to reduce their dispersal to aquatic ecosystems.

Although banned for years, organochlorine pesticides and PCBs continue to affect aquatic life, dolphins being particularly exposed. The concentrations of 31 PCB congeners, and 15 banned pesticides or metabolites were measured in 5 tissues of 68 striped dolphins stranded in the Northwestern Mediterranean coast in 2010–16. The results were compared to historical data (1988–2009) and, even though there is a slow decreasing trend, the levels in the 2010–2016 samples were still elevated based on common cetacean toxicological thresholds. A transition period in 2007–08, probably caused by a morbillivirus epizootic amplified the stranding, espacially of highly contaminated specimens. From 2010, higher proportions in parent compounds towards metabolites were observed yet again. These changing patterns were likely reflect the exposure of dolphins to the remobilization of pollutants from contaminated soils and sediments, with a prominent role of rivers. This should lead to an even slower decline of these contaminants that could last for decades, requiring new efforts to reduce their dispersal to aquatic ecosystems.

This study investigates the relationships among Ni, Cd and Pb's different chemical forms determined by different methodologies in coastal and transitional waters across a broad geographical scale. Concentrations were measured in spot samples and through passive sampling (DGT). High variability of metal concentrations was found among sampling sites and methodologies due to natural water fluctuations rather than to a given metal or method. Total dissolved metal concentrations in spot samples were lower than the EQS-WFD values. The labile fractions of Cd and Pb, measured in spot samples by Anodic Stripping Voltammetry and by DGT-ICPMS, were highly correlated. Similar labilities were found for Cd, while for Pb, the ASV labile fraction was ≈50% lower. These results reflect the pool of mobile and labile species available towards each technique kinetic window, and they seem not to be affected by discrete sampling flaws.

This study investigates the relationships among Ni, Cd and Pb's different chemical forms determined by different methodologies in coastal and transitional waters across a broad geographical scale. Concentrations were measured in spot samples and through passive sampling (DGT). High variability of metal concentrations was found among sampling sites and methodologies due to natural water fluctuations rather than to a given metal or method. Total dissolved metal concentrations in spot samples were lower than the EQS-WFD values. The labile fractions of Cd and Pb, measured in spot samples by Anodic Stripping Voltammetry and by DGT-ICPMS, were highly correlated. Similar labilities were found for Cd, while for Pb, the ASV labile fraction was ≈50% lower. These results reflect the pool of mobile and labile species available towards each technique kinetic window, and they seem not to be affected by discrete sampling flaws.