Effect of glyphosate acid on biochemical markers of periphyton exposed in outdoor mesocosms in the presence and absence of the mussel Limnoperna fortunei.

Glyphosate is currently the most widely used herbicide in agricultural production. It generally enters aquatic ecosystems through surface water runoff and aerial drift. We evaluated the effect of glyphosate acid on biochemical parameters of periphyton exposed to concentrations of 1, 3, and 6 mg/L in outdoor mesocosms in the presence and absence of the mussel Limnoperna fortunei. Periphyton ash-free dry weight, chlorophyll a content, carotene/chlorophyll a ratio, lipid peroxidation levels, and superoxide dismutase and catalase activities were determined at days 0, 1, 7, 14, and 26 of the experimental period. Ash-free dry weight was similar between control and glyphosate-treated periphyton in the absence of L. fortunei. The latter had significantly lower carotene to chlorophyll a ratios and enzyme activities, and higher lipid peroxidation levels and chlorophyll a content than the former. These results show an adverse effect of glyphosate on the metabolism of periphyton community organisms, possibly inducing oxidative stress. On the contrary, no differences were observed in any of these variables between control and glyphosate-treated periphyton in the presence of L. fortunei. Mussels probably attenuated the herbicide effects by contributing to glyphosate dissipation. The results also demonstrate that biochemical markers provide useful information that may warn of herbicide impact on periphyton communities. Environ Toxicol Chem 2017;36:1775-1784. © 2016 SETAC.

Impact of multiple anthropogenic stressors on freshwater: how do glyphosate and the invasive mussel Limnoperna fortunei affect microbial communities and water quality?

The study of the joint effect of multiple anthropogenic stressors is important because the emerging consequences are often unpredictable on the basis of knowledge of single effects. We explored the joint impact of glyphosate and the invasive golden mussel Limnoperna fortunei on freshwater phytoplankton, bacterioplankton and periphyton, and on the physical and chemical properties of the water. We manipulated both stressors simultaneously in a 25-day experiment using outdoor mesocosms; we assayed technical-grade glyphosate acid at four concentrations: 0, 1, 3 and 6 mg gly L(−1) under scenarios with and without mussels. The addition of the glyphosate significantly increased total phosphorus according to the concentration used; the high clearance rate of L. fortunei significantly decreased phytoplanktonic abundance leading to low values of turbidity. The mussel significantly stimulated the development of filamentous green algae (metaphyton). Interestingly, the combined effect revealed that L. fortunei accelerated the dissipation of glyphosate, which showed a 4-fold decrease in its half-life; this promoted the rapid bioavailability of glyphosate-derived phosphorus in the water. The interaction had a synergistic effect on soluble reactive phosphorus concentrations and was directly dependent on the concentration of glyphosate. A synergistic effect was also observed on bacterioplankton, water turbidity and metaphyton, thus inducing enhanced and rapid eutrophication. The ability of mussels to reduce glyphosate in water may be valued as positive, but our results allow us to predict that the invasion of Limnoperna fortunei in natural freshwater systems contaminated by glyphosate will accelerate the negative impact of the herbicide associated with eutrophication.

Evaluation of biochemical markers in the golden mussel Limnoperna fortunei exposed to glyphosate acid in outdoor microcosms.

In this study, the impact of technical grade glyphosate acid on Limnoperna fortunei was assessed employing outdoor microcosms treated with nominal glyphosate concentrations of 1, 3 and 6 mg L(-1). At the end of the experiment (26 days), catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST), acetylcholinesterase (AChE), carboxylesterases (CES) and alkaline phosphatase (ALP) activities, and lipid peroxidation levels were analyzed. GST and ALP activities and lipid peroxidation levels showed a significant increase with respect to controls in the mussels exposed to glyphosate (up to 90, 500 and 69 percent, respectively). CES and SOD activities showed a significant decrease in glyphosate exposed bivalves with respect to controls (up to 48 and 37 percent, respectively). CAT and AChE did not show differences between exposed and no exposed bivalves. The increase in lipid peroxidation levels and the decrease in SOD and CES activities observed in L. fortunei indicate that glyphosate had adverse effects on the metabolism of this bivalve. The results of the present study also indicate that a "multibiomarker approach" provides a more precise knowledge of the impact of glyphosate on L. fortunei.

Impact of the invasive mussel Limnoperna fortunei on glyphosate concentration in water.

The use of glyphosate has increased dramatically during the past years around the world. Microbial communities are altered when glyphosate reaches water bodies. The freshwater golden mussel Limnoperna fortunei is an invasive species that has rapidly dispersed since it was introduced in Argentina two decades ago. Mussels alter aquatic conditions through their filtrating activity by increasing water clarity and nutrient recycling. We aim to evaluate the potential capacity of the golden mussel to reduce glyphosate concentration in water, in laboratory conditions. Firstly, the evasive response of mussels to glyphosate (10, 20, and 40 mg l⁻¹) was evaluated and a toxicity test was carried out for these concentrations. A three-week experiment was then performed to assess glyphosate variation under mussel presence for two mussel sizes. Finally, mussels' role on glyphosate concentration was evaluated considering different mussel parts (living organisms and empty shells) through another three-week experiment. Laboratory experiments were performed in triplicate using 2-l microcosms. An initial glyphosate concentration between 16 and 19 mg l⁻¹ was used, and when mussels or valvae were added, 20 organisms per aquaria were used. Samples were obtained at days 0, 1, 2, 4, 8, 14, and 21. Glyphosate decreased by 40% under large mussel presence in both experiments, and was reduced by 25% in empty shell treatments. We believe that part of the herbicide that disappears from the water column is adsorbed in valvae surface, while another proportion is being mineralized by microbial communities in shells' biofilm. The mechanisms by which living mussels increase glyphosate dissipation would be degradation, possibly mediated by bacteria associated to mussel's metabolism. Glyphosate half-life depended on mussel and valvae presence and varied with mussel size. L. fortunei presence (either alive or as empty valvae) alters glyphosate concentration in water. We provide preliminary observations from laboratory experiments, with strong potential ecological consequences, about two stressors that could be acting jointly on the environment.

Direct and indirect effects of the glyphosate formulation Glifosato Atanor® on freshwater microbial communities.

Glyphosate-based formulations are among the most widely used herbicides in the world. The effect of the formulation Glifosato Atanor(®) on freshwater microbial communities (phytoplankton, bacterioplankton, periphyton and zooplankton) was assessed through a manipulative experiment using six small outdoor microcosms of small volume. Three of the microcosms were added with 3.5 mg l(-1) of glyphosate whereas the other three were left as controls without the herbicide. The treated microcosms showed a significant increase in total phosphorus, not fully explained by the glyphosate present in the Glifosato Atanor(®). Therefore, part of the phosphorus should have come from the surfactants of the formulation. The results showed significant direct and indirect effects of Glifosato Atanor(®) on the microbial communities. A single application of the herbicide caused a fast increase both in the abundance of bacterioplankton and planktonic picocyanobacteria and in chlorophyll a concentration in the water column. Although metabolic alterations related to oxidative stress were induced in the periphyton community, the herbicide favored its development, with a large contribution of filamentous algae typical of nutrient-rich systems, with shallow and calm waters. An indirect effect of the herbicide on the zooplankton was observed due to the increase in the abundance of the rotifer Lecane spp. as a consequence of the improved food availability given by picocyanobacteria and bacteria. The formulation affected directly a fraction of copepods as a target. It was concluded that the Glifosato Atanor(®) accelerates the deterioration of the water quality, especially when considering small-volume water systems.