Usefulness of oxidative stress biomarkers in native species for the biomonitoring of pesticide pollution in a shallow lake of the Austral Pampas, Argentina.

Pesticide contamination and its adverse effects on native freshwater species continue to be a worldwide major concern, mainly in developing countries. Passive biomonitoring of pesticide pollution in shallow lakes may be achieved by the simultaneous use of fish and wetland plants. Thus, the present study aimed to evaluate the occurrence of current-use pesticides in the surface water of a shallow lake of the Austral Pampas region (Buenos Aires Province, Argentina) surrounded by intensive agricultural activities and its relationship with a battery of biomarkers, including oxidative stress and genotoxicity, in two native species, the fish Oligosarcus jenynsii and the macrophyte Bidens laevis. A total of 26 pesticide residues were analyzed, and the main ones detected were glyphosate and its metabolite aminomethylphosphonic acid (AMPA), chlorpyrifos, and imidacloprid. In O. jenynsii, hydrogen peroxide (H2O2) content in the liver increased with chlorpyrifos occurrence, while malondialdehyde (MDA) levels in the brain and liver increased with the presence of both chlorpyrifos and glyphosate. In B. laevis, H2O2 and MDA levels in leaves and roots increased with AMPA occurrence. Also, leaf H2O2 contents and root MDA levels increased with chlorpyrifos concentration. In contrast, catalase and peroxidase activities in roots decreased with AMPA and chlorpyrifos occurrence. In both species, mainly H2O2 and MDA levels demonstrated their sensitivity to be used as biomarkers in the biomonitoring of current-use pesticide pollution in shallow lakes. Their use may provide information to plan strategies for environmental conservation by government institutions or decision-makers, and to assess the biota health status.

Risk evaluation and prioritization of contaminants of emerging concern and other organic micropollutants in two river basins of central Argentina.

A research gap exists in baseline concentrations of organic micropollutants in South American rivers. Identification of areas with different degrees of contamination and risk to the inhabitant biota is needed to improve management of freshwater resources. Here we inform the incidence and ecological risk assessment (ERA) of current used pesticides (CUPs), pharmaceutical and personal care products (PPCPs) and cyanotoxins (CTX) measured in two river basins from central Argentina (South America). Risk Quotients approach was used for ERA differentiating wet and dry seasons. High risk was associated to CUPs in both basins (45 % and 30 % of sites from Suquía and Ctalamochita rivers, respectively), mostly in the basins extremes. Main contributors to risk in water were insecticides and herbicides in Suquía river and insecticides and fungicides in Ctalamochita river. In Suquía river sediments, a very high risk was observed in the lower basin, mainly from AMPA contribution. Additionally, 36 % of the sites showed very high risk of PCPPs in Suquía river water, with the highest risk downstream the wastewater treatment plant of Córdoba city. Main contribution was from a psychiatric drug and analgesics. In sediments medium risk was observed at the same places with antibiotics and psychiatrics as main contributors. Few data of PPCPs are available in the Ctalamochita river. The risk in water was low, with one site (downstream Santa Rosa de Calamuchita town) presenting moderated risk caused by an antibiotic. CTX represented in general medium risk in San Roque reservoir, with San Antonio river mouth and the dam exit showing high risk during the wet season. The main contributor was microcystin-LR. Priority chemicals for monitoring or further management include two CUPs, two PPCPs, and one CTX, demonstrating a significant input of pollutants to water ecosystems from different sources and the need to include organic micropollutants in current and future monitoring.

Monensin occurrence in surface water and its impact on aquatic biota in a stream of the southeast Pampas, Argentina.

Monensin is an ionophore antibiotic used as a feed additive and growth promoter in cattle production worldwide. The occurrence of monensin in aquatic surficial ecosystems is of concern due to its possible detrimental effects on human health and native biota. Argentina is one of the most important cattle beef producers worldwide; however, there is little knowledge on the environmental occurrence of monensin and the associated risks to aquatic biota. In this study, we developed a method for the extraction and quantification of monensin in surface water; then, we evaluated the occurrence of monensin in a stream impacted by different animal husbandry's operations, and then, we analyzed the ecological implications of monensin residues on aquatic organisms using the risk quotient (RQ) method. Sampling was carried out on August 2017 from the headwaters to the floodplain of the El Pantanoso stream, Buenos Aires province, Argentina. Monensin detection frequency was 75% (n = 20). The median level was 0.40 µg/L and the maximum concentration was 4.70 µg/L. The main input of monensin was from a cattle slaughterhouse, an activity that has not been considered before in the literature as a source of emission of veterinary pharmaceuticals into the environment. The RQ assessment showed that monensin levels could have potential negative effects on aquatic biota in the sampling site closest to the cattle slaughterhouse. The data obtained in this study shows that monensin was present in El Pantanoso surface waters at levels of high ecotoxicological risk to aquatic biota.

Ecological Risk Assessment (ERA) of pesticides from freshwater ecosystems in the Pampas region of Argentina: Legacy and current use chemicals contribution.

Agricultural production in the Pampas region is one of the most important economic activities in Argentina. However, the possible environmental effects related to the growth of this activity in the last years have not been studied enough. Particularly, the effects of pesticides mixtures are a topic of great concern both for society and regulatory authorities worldwide, given the possible additive and synergistic relationships between these chemicals and their possible effects on aquatic biota. Based on a concentration addition model, this study developed an Ecological Risk Assessment (ERA) of pesticides from freshwater ecosystems in the Pampas region. For this purpose, reported pesticides concentrations available in public bibliography and a Risk Quotients (RQs) approach were used. A cumulative risk map was established to display RQs for current use pesticides (CUPs) and legacy chemicals. The ΣRQs were calculated for 66 sites, using available reported measured environmental concentrations (MECs) and predicted no effect concentrations (PNECs) of pesticides. While ΣRQ for only CUPs resulted in a high and very high risk (ΣRQ > 1) for 29% of the sites, when legacy pesticides were incorporated this percentage reached the 41% of the sites, increasing significantly the absolute values of RQ. Herbicides like glyphosate and atrazine contributed considerably to the ΣRQCUPs while organochlorines were the major contributors for ΣRQs when legacy pesticides were incorporated. Moreover, some active ingredients (acetochlor, carbendazim and fenitrothion) which are approved for their use in Argentina but banned in EU showed high contribution to ΣRQCUPs. The present study is the first attempt to develop an ERA in surface water of the Pampas region of Argentina and it provides a starting point for a more comprehensive pesticides monitoring and a further risk assessment program.

Selection of reference genes for reverse transcription-qPCR analysis in the biomonitor macrophyte Bidens laevis L.

The RT-qPCR has been the method used to analyze gene expression in plants but its benefits have not been completely exploited in the field of plants ecotoxicology when used as molecular biomarkers. The correct use of RT-qPCR demands to establish a certain number of reference genes (RG) which are expected to be invariable in their expression although it does not always happen. The main goals of this work were to: (1) analyze the stability of six potential RG, (2) establish the optimum number of RG, (3) select the most suitable RG to be applied in Bidens laevis under different test conditions and tissues and (4) confirm its convenience by normalizing the expression of one gene of interest under three different challenges. When all data were pooled together, the geNorm algorithm pointed out beta-actin and beta-tubulin (TUB) as the optimal RG pair while NormFinder algorithm selected nicotinamide adenine dinucleotide dehydrogenase (NADHD) and histone 3 (H3) as possessing the most invariable levels of expression. On the other hand, when data were grouped by tissues, ANOVA test selected H3 and TUB, while data grouped by conditions indicated that H3 and NADHD were the most stable RG under this analysis. Therefore, for a general-purpose set of RG, the overall analysis showed that a set of three RG would be optimum, and H3, TUB and NADHD were the selected ones. On the other hand, as RG can vary depending on the tissues or conditions, results achieved with ANOVA would be more reliable. Thus, appropriate normalization process would clearly need more than one RG.

Can an aquatic macrophyte bioaccumulate glyphosate? Development of a new method of glyphosate extraction in Ludwigia peploides and watershed scale validation.

Glyphosate is intensively used in agricultural fields and it is frequently detected in non-target wetland ecosystems. The floating hydrophyte Ludwigia peploides is widely distributed in American streams and it is an abundant species. Therefore, our objectives were (1) to establish and validate an extraction and quantification methodology for glyphosate in L. peploides and (2) to evaluate the role of this species as a potential glyphosate biomonitor in an agricultural watershed. We developed a new method of glyphosate extraction from leaves of L. peploides. The method recovery was 117± 20% and the matrix effect 20%. To validate the method using environmental samples, plants of L. peploides were collected in March 2016 from eight monitoring sites of El Crespo stream. Surface water and sediment samples were collected at the same time to measure glyphosate and to calculate bioconcentration factors (BCFs) and biota-sediment accumulation factors (BSAFs). Glyphosate was detected in 94.11% in leaves, the concentrations ranging between 4 and 108 µg/kg. Glyphosate was detected in surface water and sediments at 75% and 100% of the samples, at concentrations that varied between 0 and 1.7 µg/L and 5-10.50 µg/kg dry weight, respectively. The mean BCFs and BSAFs were 88.10 L/Kg and 7.61, respectively. These results indicate that L. peploides bioaccumulates glyphosate mainly bioavailable in the surface water. In this sense, L. peploides could be used as a biomonitor organism to evaluate glyphosate levels in freshwater aquatic ecosystems because, in addition to its capacity to bioconcentrate glyphosate, it is easy to sample and it has a restricted mobility.