Oxidative stress in the bivalve Diplodon chilensis under direct and dietary glyphosate-based formulation exposure.

The aim of this study was to evaluate and compare the effects on biochemical parameters and organosomatic indices in the freshwater bivalve Diplodon chilensis exposed to a glyphosate-based formulation under direct and dietary exposures (4 mg a.p./L). After 1, 7, and 14 days of exposure, reduced glutathione (GSH) and thiobarbituric acid reactive substances (TBARS) levels and the activities of glutathione-S- transferase (GST), superoxide dismutase (SOD), and catalase (CAT) were evaluated in the gills and digestive gland. The hepatosomatic (HSI) and branchiosomatic (BSI) indices were also analyzed. Direct and dietary glyphosate-based formulation exposure altered the redox homeostasis in the gills and digestive gland throughout the experimental time, inducing the detoxification response (GST), the antioxidant defenses (SOD, CAT, GSH), and causing lipid peroxidation. After 14 days of exposure, the HSI and BSI increased significantly (43% and 157%, respectively) only in the bivalves under direct exposure. Greater changes in the biochemical parameters were induced by the dietary exposure than by the direct exposure. Furthermore, the gills presented an earlier response compared to the digestive gland. These results suggested that direct and dietary exposure to a glyphosate-based formulation induced oxidative stress in the gills and digestive glands of D. chilensis. Thus, the presence of glyphosate-based formulations in aquatic ecosystems could represent a risk for filter-feeding organisms like bivalves.

The safener isoxadifen-ethyl confers fenoxaprop-p-ethyl resistance on a biotype of Echinochloa crus-galli.

BACKGROUND: Some herbicides are commercially formulated with safeners to increase crop selectivity. Fenoxaprop-p-ethyl is formulated with the safener isoxadifen-ethyl for Echinochloa crus-galli control in rice. Safeners act on crops by increasing herbicide metabolism, but this effect may also occur in weeds. The objective of this study was to investigate the effect of the safener isoxadifen-ethyl on the resistance to fenoxaprop-p-ethyl in a biotype of E. crus-galli. RESULTS: A screening of 52 biotypes identified lack of control in the biotype SANTPAT-R treated with the recommended dose of 69 g ha-1 of the commercial formulation of fenoxaprop-p-ethyl with the safener isoxadifen-ethyl. While this biotype survived doses greater than 2208 g ha-1 of the formulation fenoxaprop-p-ethyl + isoxadifen-ethyl, it was killed with 69 g ha-1 of fenoxaprop-p-ethyl without the safener. A glutathione-s-transferase (GST) enzymes inhibitor reduced the resistance factor in two dose-response curves. A minor effect of a CytP450 inhibitor was observed. The previous spraying of the safener isoxadifen-ethyl followed by fenoxaprop-p-ethyl induced survival in the resistant but not in the susceptible biotype. The GST1 and GSTF1 genes were up-regulated in the resistant biotype. ACCase gene mutations were not found, and no cross-resistance to other ACCase inhibitors was identified. CONCLUSION: The safener isoxadifen-ethyl present in the commercial herbicide formulation of fenoxaprop-p-ethyl is associated with resistance in the E. crus-galli SANTPAT-R biotype. This resistance is related with herbicide metabolization mediated by GST pathways. This is the first field-selected weed biotype with herbicide resistance due to safener presence in the sprayed formulation. © 2022 Society of Chemical Industry.

Toxicity to Rhinella arenarum tadpoles (Anura, Bufonidae) of herbicide mixtures commonly used to treat fallow containing resistant weeds: glyphosate-dicamba and glyphosate-flurochloridone.

Glyphosate (GLY)-dicamba (DIC) and GLY-flurochloridone (FLC) are herbicide mixtures which are widely used for treating fallow containing glyphosate resistant weeds. The aim of this study was to evaluate the acute toxic effects and the prevailing interactions on stage 36 tadpoles of the anuran species Rhinella arenarum when exposed to equitoxic and non-equitoxic combinations of these herbicide combinations. Experiments were realized using the following combinations of commercial formulations: 48% GLY-based Credit® + 57.71% DIC-based Banvel® and 48% GLY-based Credit® + 25% FLC-based Twin Pack Gold®. GLY-DIC and GLY-FLC equitoxic mixtures were assayed mixing each constituent with an equivalent individual toxicity able to induce the same lethality effect. After 96 h of exposure, GLY-DIC and GLY-FLC equitoxic mixtures presented toxic unit 50 values (TU50 96h) of 1.74 (confidence interval: 1.58-1.92) and 1.54 (confidence interval: 1.46-1.62) respectively, indicating the presence of a weak antagonistic interaction as TU values were greater than 1. For their part, most non-equitoxic combinations of GLY-DIC and GLY-FLC tested did not significantly differ from additivity, the only exception being when DIC and FLC were fixed at 0.33 TUs, where a weak antagonism was observed. Overall, results indicate that the toxicity of both GLY-DIC and GLY-FLC mixtures to R. arenarum tadpoles vary from additive to slightly antagonistic, depending on the proportion of constituting herbicide formulations present in the mixture.

An imazethapyr-based herbicide formulation induces genotoxic, biochemical, and individual organizational effects in Leptodactylus latinasus tadpoles (Anura: Leptodactylidae).

Genotoxic, biochemical, and individual organizational effects on Leptodactylus latinasus tadpoles were evaluated after exposure to an imazethapyr (IMZT)-based commercial herbicide formulation, Pivot® H (10.59% IMZT). A determination of the value of the lethal concentration (LC50) was determined as a toxicological endpoint. Alterations in animal behavior and morphological abnormalities as well as cholinesterase (ChE), catalase (CAT), and glutathione S-transferase (GST) activities were employed as individual sublethal endpoints. Micronuclei frequencies (MNs), binucleated cells (BNs), blebbed nuclei (BLs), lobed nuclei (LBs), notched nuclei (NTs), erythroplastids (EPs), and evaluation of DNA strand breaks were employed as genotoxic endpoints. All biomarkers were evaluated after 48 and 96 h of exposure to concentrations of IMZT within 0.07-4.89 mg/L. LC5096h values of 1.01 and 0.29 mg/L IMZT were obtained for Gosner stages 25 and 36, respectively. Irregular swimming, diamond body shape, and decreased frequency of keratodonts were detected at both sampling times. Results showed that IMZT increased GST activity and MN frequency at 48 and 96 h of exposure. Other nuclear abnormalities were also observed in the circulating erythrocytes of tadpoles, i.e., NT and BL values after 48 h, and LN, BL, and EP values after 96 h. Finally, results showed that IMZT within 0.07-0.22 mg/L increased the genetic damage index in tadpoles exposed for both exposure times (48 and 96 h). This study is the first to report the sublethal biochemical effects of IMZT in anurans and is also the first report using L. latinasus tadpoles as a bioindicator for ecotoxicological studies.

Toxic and genotoxic effects of the imazethapyr-based herbicide formulation Pivot H® on montevideo tree frog Hypsiboas pulchellus tadpoles (Anura, Hylidae).

Acute lethal and sublethal toxicity of the imidazolinone imazethapyr (IMZT)-based commercial formulation herbicide Pivot H® (10.59% IMZT) was evaluated on Hypsiboas pulchellus tadpoles. Whereas mortality was used as the end point for lethality, frequency of micronuclei (MNs) and other nuclear abnormalities as well as DNA single-strand breaks evaluated by the single cell gel electrophoresis assay were employed to test genotoxicity. Behavioral, growth, developmental, and morphological abnormalities were also employed as sublethal end points. Mortality studies revealed equivalent LC50 (96h) values of 1.49mg/L (confidence limit, 1.09-1.63) and 1.55mg/L (confidence limit, 1.51-1.60) IMZT for Gosner stage (GS) 25 and GS36, respectively. Behavioral changes, i.e., irregular swimming and immobility, as well as a decreased frequency of keratodonts were observed. The herbicide increased the frequency of MNs in circulating erythrocytes of tadpoles exposed for 48h to the highest concentration assayed (1.17mg/L). However, regardless of the concentration of the herbicide assayed, an enhanced frequency of MNs was observed in tadpoles exposed for 96h. The herbicide was able to induce other nuclear abnormalities, i.e., blebbed and notched nuclei, only when tadpoles were exposed for 96h. In addition, we observed that exposure to IMZT within the 0.39-1.17mg/L range increased the genetic damage index in treatments lasting for both 48 and 96h. This study represents the first evidence of acute lethal and sublethal effects exerted by IMZT on amphibians. Finally, our findings highlight the properties of this herbicide that jeopardize nontarget living species exposed to IMZT.

Comparative study of cytotoxic and genotoxic effects induced by herbicide S-metolachlor and its commercial formulation Twin Pack Gold® in human hepatoma (HepG2) cells.

The in vitro effects of S-metolachlor and its formulation Twin Pack Gold(®) (96% a.i.) were evaluated in human hepatoma (HepG2) cells. Cytokinesis-blocked micronucleus cytome (CBMN-cyt) and MTT assays as well as Neutral Red uptake were employed for genotoxicity and cytotoxicity evaluation. Activities were tested within the concentration range of 0.25-15 µg/ml S-metolachlor for 24h of exposure. Both compounds rendered a minor reduction in the NDI although not reaching statistical significance. Results demonstrated that the S-metolachlor was not able to induce MNs. On the other hand, 0.5-6 µg/ml Twin Pack Gold(®) increased the frequency of MNs. When cytotoxicity was estimated, S-metolachlor was not able to induce either a reduction of lysosomal or mitochondrial activity. Contrarily, whereas 1-15 µg/ml Twin Pack Gold(®) induced a significant reduction of mitochondrial activity, all tested concentrations of the formulated product induced a significant decrease of lysosomal performance as a function of the concentration of the S-metolachlor-based formulation titrated into cultures. Genotoxicity and cytotoxicity differences obtained with pure S-metolachlor and the commercial S-metolachlor-based formulation indicate that the latter may contain additional unsafe xenobiotics and support the concept of the importance of evaluating not only the active principle but also the commercial formulation when estimating the real hazard from agrochemicals.