Effects of pasture intensification and sugarcane cultivation on non-target species: A realistic evaluation in pesticide-contaminated mesocosms.

Conventional soil management in agricultural areas may expose non-target organisms living nearby to several types of contaminants. In this study, the effects of soil management in extensive pasture (EP), intensive pasture (IP), and sugarcane crops (C) were evaluated in a realistic-field-scale study. Thirteen aquatic mesocosms embedded in EP, IP, and C treatments were monitored over 392 days. The recommended management for each of the areas was simulated, such as tillage, fertilizer, pesticides (i.e. 2,4-D, fipronil) and vinasse application, and cattle pasture. To access the potential toxic effects that the different steps of soil management in these areas may cause, the cladoceran Ceriophania silvestrii was used as aquatic bioindicator, the dicot Eruca sativa as phytotoxicity bioindicator in water, and the dipteran Chironomus sancticaroli as sediment bioindicator. Generalized linear mixed models were used to identify differences between the treatments. Low concentrations of 2,4-D (<97 µg L-1) and fipronil (<0.21 µg L-1) in water were able to alter fecundity, female survival, and the intrinsic rate of population increase of C. silvestrii in IP and C treatments. Similarly, the dicot E. sativa had germination, shoot and root growth affected mainly by 2,4-D concentrations in the water. For C. sancticarolli, larval development was affected by the presence of fipronil (<402.6 ng g-1). The acidic pH (below 5) reduced the fecundity and female survival of C. silvestrii and affected the germination and growth of E. sativa. Fecundity and female survival of C. silvestrii decrease in the presence of phosphorus-containing elements. The outcomes of this study may improve our understanding of the consequences of exposure of freshwater biota to complex stressors in an environment that is rapidly and constantly changing.

Copper and cadmium, isolated and in the mixture, impact the Neotropical freshwater Calanoida copepod Notodiaptomus iheringi: A short-term approach with environmental concentrations.

Metal discharges in aquatic ecosystems are of concern since they affect different trophic levels, altering the functioning of the aquatic food chain. The metals can interact among them and with other pollutants, resulting in complex mixtures whose effects on biota are unpredictable. The impacts of copper (Cu) and cadmium (Cd), isolated and combined, on the freshwater copepod Notodiaptomus iheringi were assessed in acute and sub-chronic exposures. Species sensitivity distribution (SSD) curves were constructed for both metals. In the acute tests antagonism was observed in mortality, while in sub-chronic, mortality was not affected; however, the eggs produced and percentage of viable eggs were significantly altered. Our data suggest that egg production can be a detoxification route in N. iheringi under Cu and mixture exposure. From the SSD curves, N. iheringi was the most sensitive Brazilian species for Cu and the second most sensitive for Cd.

Environmental concentrations of cadmium and fipronil, isolated and combined, impair the survival and reproduction of a Neotropical freshwater copepod.

Anthropogenic activities such as agriculture and industry increase contaminants that reach the water bodies, potentially threatening the biota. Most likely, these pollutants occur in complex mixtures. The effects on organisms can be potentiated (synergism) or reduced (antagonism) according to the interaction between the stressors or the species. Cadmium (Cd) is a toxic metal present in phosphate fertilizers, and fipronil is an insecticide broadly used in sugarcane crops. Copepods are important energy transfer links in aquatic environments, and effects on this group impact the whole trophic chain. In this study, we evaluated the responses of the freshwater Calanoida copepod Notodiaptomus iheringi, naturally present in water bodies that can be affected by sugarcane cultures in Brazil. The organisms were exposed to environmental concentrations of Cd and fipronil, isolated and in the mixture, in acute (48 h) and sub-chronic (8 d) tests. Our data indicate that both contaminants affect the survival of the organisms in acute or sub-chronic exposures. Cadmium did not affect egg production or hatching, while fipronil impacted these endpoints negatively. The Cd-fipronil combination resulted in antagonistic responses in survival (acute and sub-chronic) and egg production. A synergistic response was observed in egg hatching. Our results suggest that Cd presents a protective effect in the mixture with fipronil; however, it is not enough to prevent egg-hatching inhibition. These responses highlight how tricky it is to deal with pollutants' interaction in environmental concentrations since synergism is the most common response to metal-pesticide mixtures. Our data point out N. iheringi as a sensitive organism in the presence of contaminants and reflects the threat of chemical mixtures in concentrations found in water bodies close to sugarcane crops in Brazil.

The acute effects of fipronil and 2,4-D, individually and in mixture: a threat to the freshwater Calanoida copepod Notodiaptomus iheringi.

The magnitude of copepods' responses to pesticides, individually and in mixture, is little understood. The aims of this study were to evaluate: (i) the effects of the pesticides fipronil and 2,4-D, individually and in mixture, on the freshwater copepod Notodiaptomus iheringi; and (ii) the survival and the feeding rate of copepods after the exposure. Acute toxicity tests using the commercial formulations of fipronil and 2,4-D, individually and in mixture, were performed. The LC10-48h, LC20-48h, and LC50-48h of fipronil to N. iheringi were 2.38 ± 0.48, 3.08 ± 1.14, and 4.97 ± 3.30 µg L-1, respectively. For 2,4-D the LC10-48h, LC20-48h, and LC50-48h were 371.18 ± 29.20, 406.93 ± 53.77, and 478.24 ± 107.77 mg L-1, respectively. Morphological damages on the copepods exposed to pesticides were observed at all concentrations. Fungal filaments covering dead organisms were presented at the treatment highest concentration (R5:7.43 ± 2.78 µg L-1 fipronil). The mixture of the pesticides presented synergistic effects on the mortality of N. iheringi. Post-exposure tests showed no difference between the treatments and the control on the mortality and on the feeding rate for 4 h. However, since delayed toxicity of pesticides can occur, longer post-exposure tests using N. iheringi should be tested. N. iheringi is a key species in the aquatic Brazilian ecosystem and showed sensitivity to fipronil and 2,4-D; thus, more studies with this species assessing other responses are recommended.

Responses of Chironomus sancticaroli to the simulation of environmental contamination by sugarcane management practices: Water and sediment toxicity.

Sugarcane management practices include the application of pesticides, including the herbicide 2,4-D and the insecticide fipronil. In addition, a by-product from the ethanol industry, called vinasse, is commonly applied to fertilize sugarcane areas. The potential risks of these practices to the edge-of-field aquatic ecosystems were assessed in the present study. This was done by contaminating mesocosms with (single and mixtures of) both pesticides and vinasse and evaluating the effects on the midge Chironomus sancticaroli through in-situ and laboratory bioassays. To this end, outdoor mesocosms were treated with fipronil (F), 2,4-D (D), and vinasse (V) alone and with the mixture of fipronil and 2,4-D (M), as well as with both pesticides and vinasse (MV). C. sancticaroli was deployed in mesocosms before contamination in cages, which were taken out 4- and 8-days-post-contamination. Water and sediment samples were also taken for laboratory bioassays on the first day of contamination, as well as 7-, 14-, 21-, 30-, 45-, and 75-days post-contamination. The responses assessed in subchronic assays (8-day) were survival, growth, head capsule width, development, and mentum deformities. Low survival occurred in the in-situ experiments of all treatments due to the low oxygen levels. In the laboratory tests, effects on survival occurred for F, V, and M over time after exposure to both water and sediment. All organisms died post-exposure to water samples from the MV treatment, even 75-days-post-contamination. Impairments in body length and head capsule width occurred for F, V, and M for water and F, V, M, and MV for sediment samples over time. All treatments increased mentum deformities in exposed larvae for any of the sampling periods. The negative effects observed were more significant in the mixture mesocosms (M and MV), thus indicating increased risks from management practices applying these compounds together or with a short time interval in crops.

Realistic exposure to fipronil, 2,4-D, vinasse and their mixtures impair larval amphibian physiology.

Expansion of sugarcane crops may have contributed to the increased contamination of native habitats in Brazil. Several species of amphibians inhabit ponds formed in flooded farmlands, where pesticide concentrations are usually high. This study evaluated the ecotoxicological effects of the sugarcane pesticides fipronil and 2,4-D, as well as the fertilizer vinasse (isolated and mixed), on physiological responses of Leptodactylus fuscus and Lithobates catesbeianus tadpoles. In situ assays were conducted in mesocosms with concentrations based on the doses recommended by the manufacturer. Vinasse (1.3% dilution) caused 100% tadpoles' mortality immediately after its application. Fipronil and/or 2,4-D altered antioxidant and biotransformation responses, induced neurotoxicity and changed lipid contents in tadpoles. A multivariate approach indicated that the mixture of pesticides induced most of the sublethal effects in both tadpole species, in addition to the isolated fipronil in L. fuscus. Fipronil alone increased glucose-6-phosphate dehydrogenase (G6PDH) activity, decreased acetylcholinesterase (AChE) and total lipid contents, and altered some individual lipid classes (e.g., free fatty acids and acetone-mobile polar lipids) in L. fuscus. The interaction between fipronil and 2,4-D in this species were more evident for lipid contents, although enzymatic alterations in G6PDH, AChE and glutathione-S-transferase (GST) were also observed. In L. catesbeianus, the mixture of pesticides reduced triglycerides and total lipids, as well as increased GST and decreased AChE activities. The detoxifying enzyme carboxylesterase was reduced by 2,4-D (alone or in mixture) in both species. Isolated pesticides also modulated specific lipid classes, suggesting their disruptive action on energy metabolism of tadpoles. Our study showed that fipronil, 2,4-D, and vinasse, individually or mixed, can be harmful to amphibians during their larval phase, causing mortality or impairing their functional responses.