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International audience; Background. Tropical coral reefs cover only ca. 0.1% of the Earth’s surface but host an outstanding biodiversity and provide important ecosystem services to millions of people living nearby. They are currently threatened by global (e.g., climate change) and local (e.g., chemical pollution) stressors that interact in different ways. While global stressors cannot be mitigated by local actions alone, local stressors can be reduced through ecosystem management. A systematic map on the impacts of chemicals arising from anthropogenic activities on tropical reef-building corals, which are the main engineer species of reef ecosystems, was published in 2021. This systematic map gathered an abundant literature (908 articles corresponding to 7937 studies), and identified four well-represented subtopics, amenable to relevant full syntheses. Here, we focused on one of the four subtopics: we aimed to systematically review the evidence on the ecotoxicological effects of chemicals on tropical reef-building corals.Methods. The evidence will be identified from the recent systematic map on the impacts of chemicals arising from anthropogenic activities on tropical reef-building corals. Especially, all studies in the map database corresponding to the knowledge cluster “evidence on the ecotoxicological effects of chemicals on corals” will be selected. To identify the evidence produced since then, a search update will be performed using a subset of the search string used for the systematic map, and titles, abstracts and full-texts will be screened according to the criteria defining the selected cluster of the map. In addition, as the eligibility criteria for the systematic review are narrower than those used to define the cluster in the systematic map, additional screening will be carried out. The included studies will then be critically appraised and a low, medium, or high risk of bias will be assigned to each study. Data will be extracted from studies and synthesised according to a strategy depending on the type of exposure and outcome. Synthesis will be mainly quantitative but also narrative, aiming to identify toxicity thresholds of chemicals for corals.

International audience; Background: Tropical coral reefs cover ca. 0.1% of the Earth's surface but host an outstanding biodiversity and provide important ecosystem services to millions of people living nearby. They are currently threatened by local stressors (e.g. nutrient enrichment and chemical pollution arising from poor land management, sewage effluents, agriculture, industry) and global stressors (mainly seawater warming and acidification, i.e. climate change). Global and local stressors interact in different ways, but the presence of one stressor often reduces the tolerance to additional stress. While global stressors cannot be mitigated solely by local actions, local stressors can be reduced through ecosystem management, therefore minimizing the impact of climate change on coral reefs. We systematically mapped the evidence of impacts of chemicals arising from anthropogenic activities on tropical reef-building corals, which are the main engineer species of reef ecosystems, to inform decision-makers on the available evidence on this topic.Methods: We searched the relevant literature using English terms combined in a tested search string in two publication databases (Scopus and Web Of Science Core Collection). The search string combined terms describing the population (tropical reef-building corals) and the exposure (chemicals). We searched for additional literature through three search engines, three dissertations repositories, 11 specialist websites, and through a call to local stakeholders. Titles, abstracts, and full-texts were successively screened using pre-defned eligibility criteria. A database of all studies included in the map with coded metadata was produced. The evidence was described and knowledge clusters and gaps were identifed through the distribution and frequency of studies into types of exposure and/or types of outcomes and/or types of study.Review findings: The initial searches identifed 23,403 articles which resulted in 15,177 articles after duplicate removal. Among them, 908 articles were retained after screening process, corresponding to 7937 studies (a study being the combination of a taxon, an exposure, and an outcome). Among these studies, 30.5% dealt with the impact of nutrient enrichment on corals while 25% concerned the impact of human activities without referenceto a chemical. The most measured outcomes were those related to the chemical concentration in corals (bioaccumulation, 25.8%), to coral physiology (16.9%), cover (14%), and mortality (9%). Half of the studies (48.4%) were experimental—the exposure was controlled by the researchers—and were conducted in laboratory conditions (39.4%) and in situ (9%). The most studied taxa, exposure, and outcomes were diferent between experimental andobservational studies.Conclusions: We identifed four well-represented subtopics that may be amenable to relevant full syntheses via systematic reviews: (1) evidence on bioaccumulation of chemicals by corals; (2) evidence on the efects of nutrient enrichment on corals; (3) evidence on the efects of human activities on corals; and (4) evidence on the ecotoxicological efects of chemicals on corals (except nutrient enrichment). The systematic map shows that corals in their natural environment can be exposed to many categories of chemicals, and that there is a complete gap in experimental research on the combined efects of more than two categories of chemicals. We therefore encourage research on this topic.

Maize cultivators often use β-triketone herbicides to prevent the growth of weeds in their fields. These herbicides target the 4-HPPD enzyme of dicotyledons. This enzyme, encoded by the hppd gene, is widespread among all living organisms including soil bacteria, which are considered as “non-target organisms” by the legislation. Within the framework of the pesticide registration process, the ecotoxicological impact of herbicides on soil microorganisms is solely based on carbon and nitrogen mineralization tests. In this study, we used more extensive approaches to assess with a lab-to-field experiment the risk of β-triketone on the abundance and the diversity of both total and hppd soil bacterial communities. Soil microcosms were exposed, under lab conditions, to 1× or 10× the recommended dose of sulcotrione or its commercial product, Decano®. Whatever the treatment applied, sulcotrione was fully dissipated from soil after 42 days post-treatment. The abundance and the diversity of both the total and the hppd bacterial communities were not affected by the herbicide treatments all along the experiment. Same measurements were led in real agronomical conditions, on three different fields located in the same area cropped with maize: one not exposed to any plant protection products, another one exposed to a series of plant protection products (PPPs) comprising mesotrione, and a last one exposed to different PPPs including mesotrione and tembotrione, two β-triketones. In this latter, the abundance of the hppd community varied over time. The diversity of the total and the hppd communities evolved over time independently from the treatment received. Only slight but significant transient effects on the abundance of the hppd community in one of the tested soil were observed. Our results showed that tested β-triketones have no visible impact toward both total and hppd soil bacteria communities.

National audience; β-triketone herbicides are among the most used herbicides in corn crop to control broadleaf weeds.These herbicides inhibit the 4-hydroxyphenylpyruvate dioxygenase (4-HPPD) and lead to bleaching anddeath of weeds. This enzyme is not only found in plants but in all living organisms, includingmicroorganisms where it takes part to the tyrosine degradation pathway. Thus, microorganismsclassified as “non-target organisms” by current EU regulation for pesticide authorization, might beimpacted by β-triketones, with possible domino effect on microbial functions supporting soilecosystem services (Thiour-Mauprivez et al. 2019). Since microorganisms have been proposed by EFSAas key-drivers to be monitored to better protect soil ecosystem services, we tested the hypothesis thathppd bacterial community can constitute a biomarker of exposure to β-triketones. Within this context,we developed a toolbox to monitor the abundance, the diversity and the activity of the hppd bacterialcommunity.Abundance and diversity of hppd bacterial community were tested in a lab-to-field experimentaldesign following the tiered-approach recommended by EFSA to conduct pesticide environmental riskassessment (ERA). Under lab conditions, soil microcosms not exposed (control) or exposed to x1 or x10times the agronomical dose of sulcotrione (active ingredient) or Decano® (one of the commercialformulation of sulcotrione) were studied. Under field conditions, samples were collected in corn cropexposed to β-triketones or not (control). Analytical chemistry was applied to all samples to study thedissipation of β-triketone, search for residues and estimate the scenario of exposure of soilmicroorganisms. Nucleic acids (DNA/RNA) were extracted from soil samples. Home-made degeneratedprimers, specific to the hppd gene of soil bacteria, allowed us to measure the abundance (quantitativePCR), the composition (α-diversity) and the diversity (β-diversity) (NGS) of the hppd bacterialcommunity (Thiour-Mauprivez et al. 2020). Finally, the inhibition of the activity of the 4-HPPD wasassessed on pure bacterial strains under wet lab conditions to measure EC50 of β-triketones.Our poster will be presented to the audience with the aim to identify the better proxy of the hppdbacterial community that could be used as a biomarker to reflect the exposure of soil microbialcommunity to β-triketone residues. As a perspective my work might be extended to other pesticidestargeting other enzymes that are also present in so call non-target organisms such as sulfonylureasinhibiting acetohydroxy acid synthase (AHAS).

National audience; The present work aimed to design a degenerate primer pair to target a large part of the hppd soil bacterial community, possibly affected by herbicides. We validated these primers by qPCR and high-throughput sequencing analysis of soil samples.

International audience; Conventional agriculture still relics on the general use of agrochemicals (herbicides, fungicides and insecticides) to control various pests (weeds, fungal pathogens and insects), to ensure the yield of crop and to feed a constantly growing population. The generalized use of pesticides in agriculture leads to the contamination of soil and other connected environmental resources. The persistence of pesticide residues in soil is identified as a major threat for in-soil living organisms that are supporting an important number of ecosystem services. Although authorities released pesticides on the market only after their careful and thorough evaluation, the risk assessment for in-soil living organisms is unsatisfactory, particularly for microorganisms for which pesticide toxicity is solely considered by one global test measuring N mineralization. Recently, European Food Safety Authority (EFSA) underlined the lack of standardized methods to assess pesticide ecotoxicological effects on soil microorganisms. Within this context, there is an obvious need to develop innovative microbial markers sensitive to pesticide exposure. Biomarkers that reveal direct effects of pesticides on microorganisms are often viewed as the panacea. Such biomarkers can only be developed for pesticides having a mode of action inhibiting a specific enzyme not only found in the targeted organisms but also in microorganisms which are considered as "non-target organisms" by current regulations. This review explores possible ways of innovation to develop such biomarkers for herbicides. We scanned the herbicide classification by considering the mode of action, the targeted enzyme and the ecotoxicological effects of each class of active substance in order to identify those that can be tracked using sensitive microbial markers.

This study presents a novel approach for assessing the risk of agrochemicals in soil microcosms through the use of non-targeted metabolomics. The metabolome of treated soils was extracted and tested through LCMS profiling in order to generate an "Environmental Metabolic Footprint" (EMF). A dynamic characterization of pollution biomarkers was obtained through a multivariate statistical analysis of EMF data, where our results show the possible evolution towards a state of resilience. The EMF methodology was applied to two β-triketone herbicides in soil microcosms: one natural, leptospermone, and one synthetic, sulcotrione. In spite of a four-fold higher application dose, leptospermone exhibited a lower resilience time than did sulcotrione (ca. 30 days vs ca. 45 days respectively).

International audience; In this study, a bacterial strain able to use sulcotrione,a β-triketone herbicide, as sole source of carbon and energy was isolated from soil samples previously treated with this herbicide. Phylogenetic study based on16S rRNA gene sequence showed that the isolate has 100 % of similarity with several Bradyrhizobium and was accordingly designated as Bradyrhizobium sp. SR1. Plasmid profiling revealed the presence of a large plasmid (>50 kb) in SR1 not cured under nonselective conditions. Its transfer to Escherichia coli by electroporation failed to induce β-triketone degrading capacity,suggesting that degrading genes possibly located on this plasmid cannot be expressed in E. coli or that they are not plasmid borne. The evaluation of the SR1 ability to degrade various synthetic (mesotrione and tembotrione) and natural (leptospermone) triketones showed that this strain was also able to degrademesotrione. Although SR1 was able to entirely dissipate both herbicides, degradation rate of sulcotrione was ten times higher than that of mesotrione, showing a greater affinity of degrading-enzyme system to sulcotrione. Degradation pathway of sulcotrione involved the formation of 2-chloro-4-mesylbenzoic acid (CMBA), previously identified in sulcotrione degradation, and of a new metabolite identified as hydroxy-sulcotrione.Mesotrione degradation pathway leads to the accumulation of-methylsulfonyl-2-nitrobenzoic acid(MNBA) and 2-amino-4 methylsulfonylbenzoic acid(AMBA), two well-known metabolites of this herbicide. Along with the dissipation of β-triketones, one could observe the decrease in 4-hydroxyphenylpyruvate dioxygenase(HPPD) inhibition, indicating that toxicity was due to parent molecules, and not to the formed metabolites. This is the first report of the isolation of bacterial strain able to transform two β-triketones.

International audience; Pseudomonas sp. 1OP, previously isolated from a French agricultural soil, has been described as the first sulcotrione degrading bacteria. Different conditions of initial pH and herbicide concentration in liquid culture were tested to evaluate the growth performances of the isolate and its degrading capacity, with sulcotrione as the sole carbon and/or energy source. Maximal growth rate (μmax) was obtained under initial neutral conditions and with initial concentration of sulcotrione close to 180 μM, and was described, during the exponential phase, by a sigmoidal curve which could be easily fitted to the modified Gompertz equation. Complementary studies carried on the CMBA by-product and on another β-triketone herbicide showed the relative specificity of the strain against sulcotrione. The sulcotrione degrading phenotype of Pseudomonas sp.1OP was shown to be lost under non-selective conditions. Plasmid-Eckardt modified method, consecutively applied for plasmid profiling, showed that this strain carries one large plasmid (>12 kb) bearing putative genes involved in sulcotrione degradation, as demonstrated by curing experiment.