Pesticide formulations contain declared active ingredients and co-formulants presented as inert and confidential compounds. We tested the endocrine disruption of co-formulants in six glyphosate-based herbicides (GBH), the most used pesticides worldwide. All co-formulants and formulations were comparably cytotoxic well below the agricultural dilution of 1% (18–2000 times for co-formulants, 8–141 times for formulations), and not the declared active ingredient glyphosate (G) alone. The endocrine-disrupting effects of all these compounds were measured on aromatase activity, a key enzyme in the balance of sex hormones, below the toxicity threshold. Aromatase activity was decreased both by the co-formulants alone (polyethoxylated tallow amine—POEA and alkyl polyglucoside—APG) and by the formulations, from concentrations 800 times lower than the agricultural dilutions; while G exerted an effect only at 1/3 of the agricultural dilution. It was demonstrated for the first time that endocrine disruption by GBH could not only be due to the declared active ingredient but also to co-formulants. These results could explain numerous in vivo results with GBHs not seen with G alone; moreover, they challenge the relevance of the acceptable daily intake (ADI) value for GBHs exposures, currently calculated from toxicity tests of the declared active ingredient alone.

A growing body of research suggests that dysbiosis of the gut microbiota induced by environmental pollutants, such as pesticides, could have a role in the development of metabolic disorders. We have examined the long-term effects of 3 doses of the Roundup(R) herbicide (made of glyphosate and formulants) on the gut microbiota in male and female Sprague-Dawley rats. A total of 141 bacteria families were identified by a 16S sequencing analysis approach. An OPLS-DA analysis revealed an increased Bacteroidetes family S24-7 and a decreased Lactobacillaceae in 8 out of the 9 females treated with 3 different doses of R (n = 3, for each dose). These effects were confirmed by repetitive sequence-based PCR fingerprinting showing a clustering of treated females. A culture-based method showed that R had a direct effect on rat gut microbiota. Cultivable species showed different sensitivities to R, including the presence of a high tolerant or resistant strain identified as Escherichia coli by 16S rRNA sequencing. The high tolerance of this E. Coli strain was explained by the absence of the EPSPS gene (coding glyphosate target enzyme) as shown by DNA amplification. Overall, these gut microbiome disturbances showed a substantial overlap with those associated with liver dysfunction in other studies. In conclusion, we revealed that an environmental concentration of R (0.1 ppb) and other two concentrations (400 ppm and 5,000 ppm) have a sex-dependent impact on rat gut microbiome composition and thus warrants further investigation.

The inheritance of historic human-induced disruption and the fierceness of its impact change aquatic ecosystems. This work reviews some of the main stressors on freshwater ecosystems, focusing on their effects, threats, risks, protection, conservation, and management elements. An overview is provided on the water protection linked to freshwater stressors: solar ultraviolet radiation, thermal pollution, nanoparticles, radioactive pollution, salinization, nutrients, sedimentation, drought, extreme floods, fragmentation, pesticides, war and terrorism, algal blooms, invasive aquatic plants, riparian vegetation, and invasive aquatic fish. Altogether, these stressors build an exceptionally composite background of stressors that are continuously changing freshwater ecosystems and diminishing or even destroying their capability to create and maintain ongoing natural healthy products and essential services to humans. Environmental and human civilization sustainability cannot exist without the proper management of freshwater ecosystems all over the planet; this specific management is impossible if the widespread studied stressors are not deeply understood structurally and functionally. Without considering each of these stressors and their synergisms, the Earth’s freshwater is doomed in terms of both quantitative and qualitative aspects.

We evaluated the effect of AsiMax 50®, a commercial formulation of 2,4-D (2,4-dichlorophenoxyacetic acid), on the structure of both micro + nano phytoplankton (>2 μm; species composition and abundance) and cytometric populations (photosynthetic picoplankton (PPP, 0.2-2 μm), which included prokaryotic phycocyanin-rich picocyanobacteria (PC-Pcy), phycoerythrin-rich picocyanobacteria (PE-Pcy) and eukaryotic phototrophs (PEuk); and bacterioplankton (HB), heterotrophic bacteria), using a microcosms-based approach and a single 7-day exposure. Assays were performed on two different microbial assemblages sampled from freshwater bodies of two contrasting turbidity status: clear (chlorophyll a = 7.6 μgL-1, turbidity = 1 NTU) and organic turbid systems (chlorophyll a = 25.0 μgL-1, turbidity = 9 NTU). For each system, the herbicide was applied to 500 mL-Erlenmeyer flasks, at seven concentration levels of the active ingredient (a.i.): 0 (control = no addition), 0.02, 0.2, 2, 20, 200 and 2,000 mg a.i.L-1. The impact of AsiMax 50® seemed to be greater in the turbid system. In this system, total abundance of living (live) micro + nano phytoplankton showed a significant increase at lower concentrations and data were fitted to a humped-shaped curve. For both clear and organic turbid systems, micro + nano phytoplankton decreased in species richness and abundance at higher herbicide concentrations. These results suggest that 2,4-D may mimic hormonal function. Some species, such as Ochromonas sp. and Chlamydomonas sp., showed different responses to herbicide exposure between water systems. In the turbid system, the increase in abundance of the PPP fraction observed at 7-d exposure was probably due to either an increase in PE-Pcy (thus suggesting the existence of auxin pathways) or a reduction in competitive pressure by micro + nano plankton. Our results provide some evidence of the importance of using community-scale approaches in ecotoxicological studies to predict changes in freshwater ecosystems exposed to a 2,4-D-based formulation. However, caution must be taken when extrapolating these effects to real scenarios, as assays were based on a laboratory microcosm experiment.