Recommended rates of azoxystrobin and tebuconazole seem to be environmentally safe but ineffective against target fungi.

The use of fungicides in agriculture has been playing a role in the enhancement of agricultural yields through the control of pathogens causing serious diseases in crops. Still, adverse environmental and human health effects resulting from its application have been reported. In this study, the possibility of readjusting the formulation of a commercial product combining azoxystrobin and tebuconazole (active ingredients - AIs; Custodia®) towards environmentally safer alternative(s) was investigated. Specifically, the sensitivity of non-target aquatic communities to each AI was first evaluated by applying the Species Sensitivity Distributions (SSDs) approach. Then, mixtures of these AIs were tested in a non-target organism (Raphidocelis subcapitata) denoting sensitivity to both AIs as assessed from SSDs. The resulting data supported the design of the last stage of this study, where mixtures of those AIs at equivalent vs. alternative ratios and rates as in the commercial formulation were tested against two target fungal species: Pyrenophora teres CBS 123929 and Rhynchosporium secalis CBS 110524. The comparison between the sensitivity of non-target aquatic species and the corresponding efficacy towards target fungi revealed that currently applied mixture and rates of these AIs are generally environmentally safe (antagonistic interaction; concentrations below the EC1 for R. subcapitata and generally below the HC5 for aquatic non-target communities), but ineffective against target organisms (maximum levels of inhibition of 70 and 50% in P. teres CBS 123929 and R. secalis CBS 110524, respectively). Results additionally suggest a potentiation of the effects of the AIs by the other formulants added to the commercial product at tested rates. Overall, this study corroborates that commercial products can be optimized during design stages based on a systematic ecotoxicological testing for ingredient interactions and actual efficacy against targets. This could be a valuable pathway to reduce environmental contamination during transition to a more sustainable agricultural production.

Ecotoxicological assessment of the herbicide Winner Top and its active substances-are the other formulants truly inert?

Formulants used in Plant Protection Products (PPPs) to promote their efficiency are normally undisclosed in the PPP documentation, unless they bear a human health or environmental hazardous potential per se. PPP regulation also demands the assessment of putative interactions among formulants within each product recipe and consequent effects, but these results are often unavailable. Such a case is that of the herbicide Winner Top (Selectis®, Portugal), which we selected as a model commercial formulation in the present study specifically aiming at (i) characterising its aquatic toxicity towards sensitive eco-receptors (Raphidocelis subcapitata, Chlorella vulgaris, Lemna minor and Lemna gibba), as well as that of its active substances (a.s.) nicosulfuron and terbuthylazine; (ii) comparing the ecotoxicity among the commercial formulation, the corresponding mixture of its a.s. and this a.s.'s mixture increasingly enriched with the formulants. Single chemical testing revealed that terbuthylazine was the strongest microalgae growth inhibitor and nicosulfuron was the strongest macrophyte growth inhibitor. On the other hand, the commercial formulation was consistently less toxic than the corresponding mixture of the a.s., suggesting that Winner Top formulants (72.9% of the commercial formulation) interact with the a.s., promoting less than additive effects in the selected non-target species. Importantly, this environmentally protective effect of the formulation can be apparent. Because macrophytes share most physiological features with the weeds targeted by the studied herbicide, it is likely that increased application doses are required to reach desired efficacy levels with the consequent detrimental increase of PPP residues load in edge-of-field freshwater ecosystems.

Overview of Chemotaxis Behavior Assays in Caenorhabditis elegans.

Environmental pollution related to anthropogenic pressures, and the associated repercussions on public health, represent a worldwide problem. Thus, the study of the effects that environmental contaminants can pose to natural ecosystems and human health is of vital importance. Laboratory model organisms such as Caenorhabditis elegans have played a significant role in clarifying multilevel effects of those agents. Although the evaluation of contaminant effects at the behavioral level of organisms is an emerging approach in ecotoxicology, studies assessing chemotaxis behavior in C. elegans within the ecotoxicological research context are still scarce. Chemotaxis studies in C. elegans have contributed to the understanding of both the neuronal mechanisms involved in the behavioral effects triggered by environmental cues and the impact of contaminants on natural ecosystems. Its compact and well-characterized nervous system, as well as the availability of transgenic strains and molecular tools, allows a detailed examination of behavioral, molecular, and genetic chemosensation mechanisms. This overview provides a summary and general comparison of methods used to measure chemotaxis behavior in C. elegans, with the aim of helping researchers select the most suitable approach in their chemotaxis studies. We compare methods based on the type of chemical tested, advantages and drawbacks of the different approaches, and specific experimental goals. Lastly, we hope to encourage the evaluation of C. elegans chemotaxis behavior in ecotoxicology studies, as well as its potential integration in standardized protocols assessing environmental quality. © 2021 Wiley Periodicals LLC.

Haloperidol Interactions with the dop-3 Receptor in Caenorhabditis elegans.

Haloperidol is a typical antipsychotic drug commonly used to treat a broad range of psychiatric disorders related to dysregulations in the neurotransmitter dopamine (DA). DA modulates important physiologic functions and perturbations in Caenorhabditis elegans (C. elegans) and, its signaling have been associated with alterations in behavioral, molecular, and morphologic properties in C. elegans. Here, we evaluated the possible involvement of dopaminergic receptors in the onset of these alterations followed by haloperidol exposure. Haloperidol increased lifespan and decreased locomotor behavior (basal slowing response, BSR, and locomotion speed via forward speed) of the worms. Moreover, locomotion speed recovered to basal conditions upon haloperidol withdrawal. Haloperidol also decreased DA levels, but it did not alter neither dop-1, dop-2, and dop-3 gene expression, nor CEP dopaminergic neurons' morphology. These effects are likely due to haloperidol's antagonism of the D2-type DA receptor, dop-3. Furthermore, this antagonism appears to affect mechanistic pathways involved in the modulation and signaling of neurotransmitters such as octopamine, acetylcholine, and GABA, which may underlie at least in part haloperidol's effects. These pathways are conserved in vertebrates and have been implicated in a range of disorders. Our novel findings demonstrate that the dop-3 receptor plays an important role in the effects of haloperidol.

Measurement of the Effects of Metals on Taxis-to-Food Behavior in Caenorhabditis elegans.

Chemosensation in nematodes is linked to processes that affect their ability to survive, such as the search for food and the avoidance of toxic substances. Since the 1970s, numerous studies have assessed chemotaxis in the nematode species Caenorhabditis elegans, focusing on a multitude of agents, including bacteria (food), ions, salts, hormones, volatile organic compounds, and, to a lesser extent, metal-contaminated medium/food. The few studies evaluating metal exposure have reported a variety of responses (neutral, attraction, avoidance), which generally appear to be contaminant and/or concentration specific. Differences in experimental designs, however, hinder appropriate comparison of the findings and attainment of firm conclusions. Therefore, we herein propose and describe a detailed protocol for the assessment of the effects of metals on taxis-to-food behavior in C. elegans. Distinct approaches are proposed in two innovative stages of testing to (1) screen metals' effects on taxis-to-food behavior and (2) classify the behavioral response as attraction/avoidance/indifference or preference. Use of such a standard protocol will allow for easy comparison across studies and direct interpretation of results. Findings using this model system can contribute to a deeper understanding of the real risks of metal contamination to nematodes and how such contaminants could impact ecosystems in general, given the key environmental roles that these organisms play. © 2021 Wiley Periodicals LLC. Basic Protocol: Assessing the effects of metal contamination on taxis-to-food behavior in Caenorhabditis elegans Support Protocol 1: Synchronization of C. elegans by hand-picking gravid worms Support Protocol 2: Synchronization of C. elegans by using a bleaching solution.

Manganese in the Diet: Bioaccessibility, Adequate Intake, and Neurotoxicological Effects.

Manganese (Mn) is an essential element that participates in several biological processes. Mn serves as a cofactor for several enzymes, such as glutamine synthetase and oxidoreductases, that have an important role in the defense of the organisms against oxidative stress. The diet is the main source of Mn intake for humans, and adequate daily intake levels for this metal change with age. Moreover, in higher amounts, Mn may be toxic, mainly to the brain. Here, we provide an overview of Mn occurrence in food, addressing its bioaccessibility and discussing the dietary standard and recommended intake of Mn consumption. In addition, we review some mechanisms underlying Mn-induced neurotoxicity.

Mixture toxicity assisting the design of eco-friendlier plant protection products: a case-study using a commercial herbicide combining nicosulfuron and terbuthylazine.

The development of environmentally friendly plant protection products (PPPs), including pesticides, is a challenge nowadays. A commercial herbicide combining terbuthylazine and nicosulfuron as active substances (a.s.) was selected as a model PPP. The suitability of manipulating the ratio between a.s. towards alternative formulations with reduced impacts in a non-target indicator (Lemna minor) was assessed. The efficacy of such eco-friendlier a.s. ratios was then assessed using a target weed, Portulaca oleracea. Single and mixture toxicity testing with L. minor revealed an antagonistic joint action of the a.s., suggesting an environmentally protective effect of the combination compared to single dosing of a.s. The efficacy testing against the target weed of single and combined treatments of the a.s. showed that (i) the a.s. behave antagonistically throughout the whole P. oleracea response surface; (ii) there were no environmentally safe a.s. combinations ensuring target-efficacy; (iii) terbuthylazine alone was effective in controlling P. oleracea with no environmental hazardous potential, dosed at concentrations 10-fold lower than those involved in commercially recommended application doses. Overall, this case-study suggests that modelling tools widely used in the field of environmental risk assessment of PPPs may also have application in PPP design stages for a more efficient meeting of efficacy and environmental friendliness requirements.