Effects of herbicides and fertilization on biofilms of Pampean lotic systems: A microcosm study.

We experimentally assessed the impact of the application of herbicides and fertilizers derived from agricultural activity through the individual and simultaneous addition of glyphosate, atrazine, and nutrients (nitrogen 'N' and phosphorus 'P') on the biofilm community and their resilience when the experimental factors were removed. We hypothesize that i) the presence of agrochemicals negatively affects the biofilm community leading to the simplification of the community structure; ii) the individual or simultaneous addition of herbicides and nutrients produces differential responses in the biofilm; and iii) the degree of biofilm recovery differs according to the treatment applied. Environmentally relevant concentrations of glyphosate (0.7 mgL-1), atrazine (44 µgL-1), phosphorus (1 mg P L-1 [KH2PO4]), and nitrogen (3 mg N L-1[NaNO3]) were used. Chlorophyll a, ash-free dry weight, abundance of main biofilm groups and nutrient contents in biofilm were analyzed. At initial exposure time, all treatments were dominated by Cyanobacteria; through the exposure period, it was observed a progressive replacement by Bacillariophyceae. This replacement occurred on day 3 for the control and was differentially delayed in all herbicides and/or nutrient treatments in which the abundance of cyanobacteria remains significant yet in T5. A significant correlation was observed between the abundance of cyanobacteria and the concentration of atrazine, suggesting that this group is less sensitive than diatoms. The presence of agrochemicals exerted differential effects on the different algal groups. Herbicides contributed to phosphorus and nitrogen inputs. The most frequently observed interactions between experimental factors (nutrients and herbicides) was additivity excepting for species richness (antagonistic effect). In the final recovery time, no significant differences were found between the treatments and the control in most of the evaluated parameters, evincing the resilience of the community.

Spatial distribution of pesticides in surface water of the Estacas stream (Argentine Espinal region) associated with crop production.

The Espinal region (Entre Ríos, Argentina) has suffered land use changes caused by expansion of the agricultural frontier. This expansion has led to an increased use of pesticides. This study is aimed at better understanding the spatial distribution of pesticides in surface water of the Estacas stream, a representative basin of the Espinal region, associated with crop production. The location and proportion of area with soybean, maize, and wheat crops in each catchment area of the basin were estimated, and surface water samples were taken to perform a pesticide screening during a period of one year. Soybean represented approximately 71% of the total cultivated area of the basin, whereas maize and wheat accounted for 15% and 14%, respectively. The analysis of 125 analytes showed the presence of 19 pesticides. The pesticide load maps showed that atrazine was detected in an area of relatively low catchment compared to other pesticides as glyphosate, which is applied in all the agricultural fields of the basin. The load of metolachlor and S-metolachlor covered a large area of the basin. The highest recorded concentrations of these pesticides were 86 µg L-1 of atrazine, 24 µg L-1 of metolachlor, 19 µg L-1 of glyphosate, and 15 µg L-1 of S-metolachlor. The results allow better understanding the environmental distribution of pesticides associated with pest control in the crops of the basin studied, the doses and times of application, and the variation in the rainfall in the basin. This study provides relevant information about how aquatic ecosystems in agricultural basins receive the diffuse contribution of pesticides, representing potential sources of water pollution. Also, the results allow supporting the design of agricultural practices and politics to improve land-use planning for the development of sustainable basins.

Bioaccumulation in freshwater crabs. Endosulfan accumulation in different tissues of Zilchiopsis collastinensis P. (Decapoda: Trichodactylidae).

We examined the bioaccumulation of α- and ß-endosulfan and endosulfan sulfate in tissues from the crab Zilchiopsis collastinensis. There was more endosulfan accumulated in the hepatopancreas (from <2 to 467.8 ng g(-1)) than in the gonads (from <2 to 52.1 ng g(-1)) or muscles (<2 ng g(-1)). The endosulfan concentrations in the hepatopancreas decreased over time and with the endosulfan dilution (p < 0.05). In the gonads there was little bioaccumulation, which did not vary over time (p > 0.05). The hepatopancreas is a dynamic organ that is able to depurate itself, whereas the gonads act as a sink for pesticides.

Bioaccumulation of abacavir and efavirenz in Rhinella arenarum tadpoles after exposure to environmentally relevant concentrations.

Antiretrovirals are pharmaceuticals used in the treatment of the human immunodeficiency virus; they are contaminants of emerging concern that have received considerable attention in recent decades due to their potential negative environmental effects. Data on the bioaccumulation and possible environmental risks posed by these drugs to aquatic organisms are very scarce. Therefore, the aim of this study was to evaluate the bioaccumulation of abacavir and efavirenz in Rhinella arenarum tadpoles subjected to acute static toxicity tests (96 h) at environmentally relevant concentrations. The analytical procedure consisted of the development and optimization of a method involving ultra-high performance liquid chromatography with tandem mass spectrometry detection. The instrumental conditions, optimized by design of experiments using the response surface methodology, yielded limits of detection of 0.3 µg L-1 for abacavir and 0.9 µg L-1 for efavirenz; and limits of quantification of 1.9 µg L-1 for abacavir and 5.6 µg L-1 for efavirenz. Subsequently, the bioaccumulation of the pharmaceutical drugs in tadpoles was evaluated at three exposure concentrations. Efavirenz displayed the highest bioaccumulation levels. This study shows the bioaccumulation potential of abacavir and efavirenz in amphibian tadpoles at exposure concentrations similar to those already detected in the environment, indicating an ecological risk for R. arenarum and probably other aquatic organisms exposed to these drugs in water bodies.

Influence of abiotic factors (water activity and temperature) on growth and aflatoxin production by Aspergillus flavus in a chickpea-based medium.

The effect of water activity (aW; 0.87, 0.90, 0.92, 0.94, 0.96, 0.98 and 0.99), temperature (15, 25, and 30 °C), incubation time (5, 10, 14, and 21 days), and their interactions on mycelial growth and aflatoxin production in a chickpea-based medium by three Aspergillus flavus strains isolated from chickpea grains in Argentina was evaluated. Maximum growth rates were obtained at the highest aW (0.99) and 30 °C, with growth decreasing as the aW of the medium was reduced. Maximum levels of aflatoxins were produced at 0.99 aW and 25 °C after 5 days of incubation for two strains, and at 25 °C and 0.96 aW after 21 days of incubation for the third strain. The aflatoxin concentrations varied considerably depending on the aW and temperature interactions assayed. Two-dimensional profiles of aW by temperature interactions were developed from these data to identify areas where conditions indicate a significant risk from aflatoxin accumulation on chickpea. This study provides useful baseline data on conditions representing a high and a low risk for contamination of chickpea by aflatoxins which is of greater concern because this pulse is destined mainly for human consumption.

Quality of the surface water of a basin affected by the expansion of the agricultural frontier over the native forest in the Argentine Espinal region.

Land use changes have led to the degradation of multiple ecosystem services and affected the quality of aquatic ecosystems. The aims of this study were (i) to assess the expansion of the agricultural border over the native forest of an Argentinean stream basin and (ii) to characterize the surface water quality, considering physicochemical parameters, and pesticide concentrations. The agricultural frontier expansion was estimated through the analysis of satellite image coverage. Samples of surface water were taken bimonthly for 2 years. The native forest cover decreased from 72% in 1987 to 60% in 2017 due to the sustained increase in agricultural activities. In surface water, the concentrations of cations decreased: Na > Ca > K > Mg, whereas those of anions decreased: HCO3 > > Cl > SO4 > PO4. The 84 surface water samples analyzed revealed 25 pesticides, including herbicides (44%), insecticides (28%), and fungicides (28%). Herbicides were detected in more than 60% of the samples. 2,4-D, atrazine, cyproconazole, diazinon, glyphosate, AMPA, and metolachlor were detected in all the study sites and sometimes, 2,4-D, atrazine, dicamba, and metolachlor concentrations exceeded the guideline levels. The high sampling frequency of this study and the two annual cycles of crops in the basin enabled sensing of pesticide molecules and concentrations that had not been previously detected, indicating diffuse contamination. These findings signal an emergent challenge on the Espinal agro-ecosystem integrity due to changes in land use.

Validation of a high-throughput method for analysis of pesticide residues in hemp and hemp products.

Hemp has been an agricultural commodity for millennia, and it has been undergoing a resurgence in interest and production due to its high content of cannabinoids, protein, fiber and other ingredients. For legal possession and use throughout the USA, hemp and hemp products must have delta-9-tetrahydrocannabinol (THC) concentration < 0.3%. As with most crops, pesticides may be applied when farming hemp, which need to be monitored in food, feed, and medicinal products. The aim of this work was to evaluate and validate the recently developed "quick, easy, cheap, effective, rugged, safe, efficient, and robust" (QuEChERSER) sample preparation mega-method to determine pesticide residues in hemp plants, flowers, powders, oils, and pellets. High-throughput analysis of final extracts for 106 targeted pesticides and metabolites from North American monitoring lists entailed: 1) ultrahigh-performance liquid chromatography - tandem mass spectrometry (UHPLC-MS/MS) with column back-flushing, and 2) instrument-top sample preparation + low-pressure gas chromatography (ITSP+LPGC-MS/MS). In QuEChERSER, 2 g sample is extracted with 10 mL 4/1 (v/v) acetonitrile/water by mechanical shaking for 10 min, followed by 3 min centrifugation. For LC, 0.2 mL of extract is taken and solvent exchanged into initial mobile phase followed by 5 min ultra-centrifugation prior to the 10 min analysis. For GC-amenable pesticides, the remaining initial extract is partitioned with 4/1 (w/w) anh. MgSO4/NaCl, and 1 mL is taken for automated ITSP cleanup in parallel with 10 min LPGC analysis. In the former case, the UHPLC column is back-flushed with 1/1 (v/v) methanol/acetonitrile for 3 min between each injection to keep the system clean and avoid ghost peaks. Multi-level, multi-day validation results achieved 70-120% recoveries with RSDs < 20% for more than 80% of the analytes in hemp protein powder, oil, pellets, and fresh plant (dried hemp plant and flower were too complex). Limits of quantification (LOQs) were < 10 ng/g were achieved for nearly all pesticides, yielding 2.8% false negatives among >13,000 analyte results in the spiked samples. The QuEChERSER method was demonstrated to meet the challenge for several complex hemp matrices.

Influence of rainfall and seasonal crop practices on nutrient and pesticide runoff from soybean dominated agricultural areas in Pampean streams, Argentina.

An increase in the spatial variability of rainfall is expected due to climate change. This implies increasing rainfall rates during spring and summer in the Pampas region, Argentina, period of maximum application of agrochemicals, which might cause an increase in pesticides and nutrients carried to surface water systems, as runoff by rainfall is one of the main pathways for diffuse pollution. The crops phenological stage can also affect pesticide and nutrient runoff since the applied agrochemicals and soil cover differ in each stage. In this study, we assessed the influence of rainfall and seasonal crop practices on water quality (nutrient and pesticide concentrations) in three streams in the Pampas region, Argentina. Five sampling campaigns were performed before and after three rainfall events during two different seasons of crop practices (SCP1, SCP2) and the physicochemical characteristics of the stream and runoff water were analyzed. The pesticide concentrations in the streams presented a general increase immediately after the rainfall event. Water quality was also affected, as an increase in ammonium, soluble reactive phosphorus (SRP), biological oxygen demand (BOD), and turbidity was observed. The crops phenological stage influenced pesticide and nutrient types and concentrations detected in the streams. During SCP1, mainly characterized by chemical fallow and sowing of soybean and vegetative growth and flowering of corn, ammonium, SRP, BOD, turbidity, and some pesticides, such as metolachlor, showed significantly higher results than those found in SCP2 (grain filling and vegetative growth of soybean and corn sowing). The pesticide concentrations detected in runoff water depended mostly on the pesticide solubility, the lateral slope of the streams, and the percentage of woody riparian vegetation cover. The results obtained show the relevance of assessing the influence of rainfall and crops phenological stages on the dynamics of surface water and on pesticide and nutrient runoff for environmental monitoring.

Effects of the emulsifiable herbicide Dicamba on amphibian tadpoles: an underestimated toxicity risk?

The effects of exposure to the herbicide Dicamba (DIC) on tadpoles of two amphibian species, Scinax nasicus and Elachistocleis bicolor, were assessed. Mortality and biochemical sublethal effects were evaluated using acetylcholinesterase (AChE), glutathione S-transferase (GST), glutathione reductase (GR), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) activities and thyroid hormone (T4) levels. The LC50 value at 48h was 0.859 mg L-1 for S. nasicus and 0.221 mg L-1 for E. bicolor tadpoles. After exposure to sublethal DIC concentrations for 48 h, GST activity increased in S. nasicus but significantly decreased in E. bicolor with respect to controls. GR activity decreased only in S. nasicus at all the tested DIC concentrations. AChE activity was significantly inhibited in both S. nasicus and E. bicolor tadpoles at 48 h. DIC also caused significant changes in transamination, as evidenced by an increase in AST and ALT activities in both amphibian species. T4 levels were higher in DIC-treated tadpoles of both species than in controls. The DIC-induced biochemical alterations in glutathione system enzymes and transaminases indicate lesions in liver tissues and cellular function. Moreover, the observed AChE inhibition could lead to the accumulation of acetylcholine, excessively stimulating postsynaptic receptors, and the increase in T4 levels in both species may indicate an overactive thyroid. The commercial DIC formulation showed a high biotoxicity in the two amphibian native species after short-term exposure, controversially differing from the toxicity level indicated in the official fact sheet data. This fact highlights the need for an urgent re-categorization and reevaluation of DIC toxicity in native species.

Determination of glyphosate, AMPA and glufosinate in dairy farm water from Argentina using a simplified UHPLC-MS/MS method.

Argentina, together with the USA and Brazil, produces approximately 80% of the total worldwide glyphosate loadings. The development of a simplified ultra-high performance liquid chromatographic tandem mass spectrometric method (UHPLC-MS/MS) for the determination of glyphosate, aminomethylphosphonic acid (AMPA) and glufosinate in water is described, including studies of several alternatives of 9-fluorenylmethylchloroformate (FMOC-Cl) derivatization and pretreatment steps. The proposed method includes acidification and neutralization of a low sample volume (3 mL), 2 hours derivatization step, cleanup with dichloromethane, followed by reverse phase UHPLC-MS/MS determination of the analytes. Figures of merit were satisfactory in terms of linearity, selectivity, accuracy and intermediate precision (%REC 70-105% with RSD < 15%). Limits of quantification (LOQ) were suitable for monitoring purposes (0.6, 0.2, 0.1 µg/L for glyphosate, AMPA and glufosinate respectively). The validated methodology was applied for the analysis of livestock wells waters from 40 dairy farms located in the central region of Argentina. Glyphosate and AMPA were quantified in 15% and 53% of the analyzed samples with concentrations ranging from 0.6-11.3 µg/L and 0.2-6.5 µg/L respectively. Greater concentrations of glyphosate were also verified in waters from open-reservoir tanks, which are directly exposed to the farm environment. In these cases glyphosate and AMPA occurrence increased, being quantified in the 33% and 61% of the samples with values ranging 0.6-21.2 µg/L and 0.2-4.2 µg/L respectively. Also in this case glufosinate was found in 52% samples at

Glyphosate loss by runoff and its relationship with phosphorus fertilization.

The aim of this study was to evaluate the relationship between glyphosate and phosphate fertilizer application and their contribution to surface water runoff contamination. The study was performed in Aquic Argiudoll soil (Tezanos Pinto series). Four treatments were assessed on three dates of rainfall simulation after fertilizer and herbicide application. The soluble phosphorus in runoff water was determined by a colorimetric method. For the determination of glyphosate and aminomethylphosphonic acid (AMPA), a method based on fluorenylmethyloxycarbonyl (FMOC) group derivatization, solid phase extraction (SPE) purification, and ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was employed. The application of phosphorus fertilizer resulted in an increased loss of glyphosate by runoff after 1 day of application. These results suggest the need for further study to understand the interactions and to determine appropriate application timing with the goal of reducing the pollution risk by runoff.