Development of new approach methodologies is urgently needed to characterize the likelihood that complex mixtures of chemicals affect water quality. Omics advances in ecotoxicology allow assessment on a broadest coverage of disrupted biological pathway by mixtures. Here the usefulness of transcriptomic analyses for evaluation of combined effects and identification of main effect components are explored. Two artificial mixtures (Mix 1 and Mix 2) were tested by a concentration-dependent reduced zebrafish transcriptome (CRZT) approach and toxicity bioassays using zebrafish embryos. Then, the toxicities and transcriptomic effects of 12 component chemicals on embryos were incorporated into additivity models to characterize the combined effects of chemicals in mixtures and to identify the main bioactive compounds. Mix 1 and Mix 2 displayed similar embryo toxicities (LD₅₀: 6.6 μM and 8.7 μM, respectively), however, Mix 2 elicited broader biological process perturbations and 5-fold higher transcriptome potency (point of departure eliciting a 20% pathway response, PODₚₐₜₕ₂₀) than Mix 1. The predicted mixture toxicities derived from additivity expectations deviated by 2-fold or less from the measured embryo toxicities except for the Jaw defect endpoint; most biological processes deviated by 3-fold or less. Finally, diclofenac (DFC) and propiconazole (PCZ) were identified as the main contributing components (≥80% explanation) to the embryo toxicity and biological process perturbations by Mix 1. While DFC and chlorophene (CLP) explained up to 80% of the embryo toxicities and biological effects of Mix 2 associated with development and Metabolism processes. The CRZT approach provides a powerful tool for assessment of biological pathways perturbed by chemicals in mixtures and for identification of main bioactive compounds.

Carbendazim is a fungicide commonly used as active substance in plant protection products and biocidal products, for instance to protect facades of buildings against fungi. However, the subsequent occurrence of this fungicide and potential endocrine disruptor in the aqueous environment is a major concern. In this study, high resolution mass spectrometry shows that carbendazim can be detected with an increasing abundance from the source to the mouth of the River Rhine. Unexpectedly, the abundance of carbendazim correlates poorly with that of other fungicides used as active ingredients in plant protection products (r² of 0.32 for cyproconazole and r² of 0.57 for propiconazole) but it correlates linearly with that of pharmaceuticals (r² of 0.86 for carbamazepine and r² of 0.89 for lamotrigine). These results suggest that the occurrence of carbendazim in surface water comes mainly from the discharge of treated domestic wastewater. This hypothesis is further confirmed by the detection of carbendazim in wastewater effluents (n = 22). In fact, bench-scale leaching tests of textiles and papers revealed that these materials commonly found in households could be a source of carbendazim in domestic wastewater. Moreover, additional river samples collected nearby two paper industries indicate that the discharge of their treated process effluents is also a source of carbendazim in the environment. While characterizing paper and textile as overlooked sources of carbendazim, this study also shows the biocide as a possible ubiquitous wastewater contaminant that would require further systematic and worldwide monitoring due to its toxicological properties.

The Red River Delta is a major agricultural production area of Vietnam with year-round use of pesticides for paddy rice cultivation and other production systems. The delta is protected from flooding, storm surges and saline water intrusion by a sophisticated river and sea-dyke system. Little is known about the effects of such a dyke system on pesticide pollution in the enclosed landscape. Our aim was to address this gap by i) determining pesticide prevalence in soils and sediments within a dyked agricultural area, and by ii) assessing whether and to which degree this dyke system might affect the spatial distribution of pesticides. After sampling paddy rice fields (topsoil) and irrigation ditches (sediment) perpendicular to the dyke in Giao Thuy district, we analysed 12 of the most commonly used pesticides in this area. In soils, we detected most frequently isoprothiolane (100% detection frequency), chlorpyrifos (85%) and propiconazole (41%) while in sediments isoprothiolane (71%) and propiconazole (71%) were most frequently found. Maximum concentrations reached 42.6 μg isoprotiolane kg⁻¹ in soil, and 35.1 μg azoxystrobin kg⁻¹ in sediment. Our results supported the assumption that the dyke system influenced residue distribution of selected pesticides. More polar substances increasingly accumulated in fields closer to the sea-dyke (R² = 0.92 for chlorpyrifos and 0.51 for isoprothiolane). We can thus support initiatives from local authorities to use the distance to dykes as a mean for deliniating zones of different environmental pollution; yet, the degree at which dykes influence pesticide accumulation appear to be compound specific.

Fungicides are used to prevent foliar diseases on a wide range of vegetable, field, fruit, and ornamental crops. They are generally more effective as protective rather than curative treatments, and hence tend to be applied before infections take place. Less than 1% of US soybeans were treated with a fungicide in 2002 but by 2006, 4% were treated. Like other pesticides, fungicides can move-off of fields after application and subsequently contaminate surface water, groundwater, and associated sediments. Due to the constant pressure from fungal diseases such as the recent Asian soybean rust outbreak, and the always-present desire to increase crop yields, there is the potential for a significant increase in the amount of fungicides used on US farms. Increased fungicide use could lead to increased environmental concentrations of these compounds. This study documents the occurrence of fungicides in select US streams soon after the first documentation of soybean rust in the US and prior to the corresponding increase in fungicide use to treat this problem. Water samples were collected from 29 streams in 13 states in 2005 and/or 2006, and analyzed for 12 target fungicides. Nine of the 12 fungicides were detected in at least one stream sample and at least one fungicide was detected in 20 of 29 streams. At least one fungicide was detected in 56% of the 103 samples, as many as five fungicides were detected in an individual sample, and mixtures of fungicides were common. Azoxystrobin was detected most frequently (45% of 103 samples) followed by metalaxyl (27%), propiconazole (17%), myclobutanil (9%), and tebuconazole (6%). Fungicide detections ranged from 0.002 to 1.15 μg/L. There was indication of a seasonal pattern to fungicide occurrence, with detections more common and concentrations higher in late summer and early fall than in spring. At a few sites, fungicides were detected in all samples collected suggesting the potential for season-long occurrence in some streams. Fungicide occurrence appears to be related to fungicide use in the associated drainage basins; however, current use information is generally lacking and more detailed occurrence data are needed to accurately quantify such a relation. Maximum concentrations of fungicides were typically one or more orders of magnitude less than current toxicity estimates for freshwater aquatic organisms or humans; however, gaps in current toxicological understandings of the effects of fungicides in the environment limit these interpretations.

Eroded soil material may be an important transporting agent for pesticides that are strongly sorbed to soil. The abilityof the fungicide propiconazole to interact with colloidal andparticulate materials has been studied by means of sorptionand desorption experiments. Size separation of silty clay soilfrom Mørdre, Norway and subsequent characterization showedthat different size fractions of soil possessed different physical and chemical properties and, therefore, different capacity to associate with propiconazole. A large part of the soil organic carbon was associated with coarser material (2–0.02 mm), which also showed higher affinity towards propiconazole than for smaller size fractions (<20 and <2 μm). Similar K ₒc values (2306 and 2244) for the size fractions <2 and <20 μm indicate that organic carbon played a dominant role in the sorption of propiconazole. Furthermore, organic carbon associated with these size fractions seemed to have similar properties withrespect to binding of propiconazole. Although, poor in organiccarbon (0.4%), the smallest size fraction (<2 μm) had higher sorption capacity for propiconazole compared to the medium size fraction (<20 μm). Higher sorption for the smallest size fraction (<2 μm) is probably due to higherspecific surface area, cation exchange capacity and content of Fe/Al oxides (free, organically bound and amorphous oxides) than the other size fractions. Results from the desorption experiments indicate that a part of propiconazole associates with sites in the soil material that resist desorption. Fluvialsediments originating from propiconazole treated fields may, therefore, represent potential reservoirs of propiconazole.Treatment with H₂O₂ modified the sorption/desorptioncharacteristics of the soil beyond that which could be expectedsimply by the removal of organic material. The pH values for all the size fractions decreased, and the specific surface areaof the medium sized fraction (<20 μm) increased from 14 to 19 m² g⁻¹ after the treatment with H₂O₂,probably due to disruption of the aggregate structure. Carrying out fractionation and separation procedures, it is important to be aware of physical and chemical changes that areintroduced during the different steps. An effort should be made to develop fractionation methods that keep the original characteristics of the soil material as intact as possible.

Brazil is one of the largest consumers of pesticides in the world, and these chemicals present a high contamination risk for the country’s water bodies. The mechanisms of mobilization and transport of pesticides from cropland to river systems are controlled by runoff and erosion processes occurring at the catchment scale. In addition to the excessive use of pesticides, the transport processes of these substances are also accelerated by inadequate soil management and the absence of soil conservation measures at the catchment scale. The current research relied on hydrological monitoring to investigate the transport and persistence of pesticides in response to hydrological dynamics. The study was conducted in the Conceição River watershed where runoff and suspended sediment fluxes are continuously monitored at the outlet. This study area is representative of the grain production system in southern Brazil including the application of large amounts of pesticides combined with extensive runoff and erosion problems. Sample collection in the river for pesticide analysis included the analysis of both water and suspended sediment. The sediment deposit analysis was performed in a single location at 4 depths. Results demonstrate the occurrence of pesticides including simazine, 2,4-D, carbendazim, imidacloprid, tebuconazole, propiconazole, tetraconazole, and trifloxystrobin in water, while glyphosate and AMPA were detected in suspended sediments, and AMPA and carbendazim were found in sediment deposits. The study demonstrated the strong dependence of the mechanisms of pesticide mobilization and transport in the catchment with the intra- and interevent variability of hydro-sedimentary processes. Pesticide detections can be related to several factors, including the magnitude of the rainfall event, the period of pesticide application, or the transport of suspended sediment. All these factors are correlated, and the mechanisms of transportation play an important role in the connections between sink and sources. The results suggest that pesticide monitoring should take into account the runoff and erosion pathways in each particular catchment, and it should especially include the monitoring of major rainfall events for identifying and quantifying the occurrence of pesticides in the environment. The transport of pesticides indicates to be potentiated by intensive pesticide use, the magnitude of rainfall-runoff events, and the absence of runoff control measures (e.g., terracing). These results demonstrate that water and soil conservation techniques should be planned and coordinated at the watershed scale to reduce the connectivity of water and sediment flows from agricultural areas to river systems with the implementation of effective runoff control practices. This will control the mobilization agents (runoff), as well as limit the connection between the sources and the water bodies.

Many of South Africa’s freshwater impoundments are compromised by pesticide pollution, and the Roodeplaat Dam, near Pretoria, is no exception. This paper is, to the best of our knowledge, the first report of the herbicides Dacthal, metribuzin, simazine, tebuthiuron, terbuthylazine, and the fungicides azoxystrobin, carbendazim, epoxiconazole, metalaxyl (Ridomil), propiconazole, pyrimethanil and thiabendazole in a South African freshwater impoundment. This short note reports on the screening results of water and muscle tissue samples against a comprehensive library of pesticides, herbicides and fungicides in the polluted Roodeplaat Dam. Muscle samples of Oreochromis mossambicus screened positive for p,p'-DDE and p,p'-DDD and for DCPA (chlorthal-methyl). The muscle tissue of Clarias gariepinus screened positive for p,p'-DDE and p,p'-DDD, chlorpyrifos, trans-chlordane, DCPA and terbuthylazine. The presence of these pesticides, herbicides and fungicides in this impoundment is of great concern as there is substantial evidence of adverse health effects in fish exposed to these chemicals.

Propiconazole (PCZ) is a widely used fungicide around the world and was frequently detected in surface waters, which would pose risk to aquatic organisms. Previous studies indicated that PCZ has high toxicity to different kinds of fish. However, most of the studies focus on the toxicity and mechanisms of PCZ to adult fish, the potential toxicity mechanism of PCZ to fish embryos is still poorly understood. The present study investigated the effects of PCZ on content of reactive oxygen species (ROS) and malondialdehyde (MDA); activities of superoxide dismutase (SOD), catalase (CAT), and Na⁺-K⁺-ATPase; and expression level of genes related to oxidative stress, cell apoptosis, and innate immune system in zebrafish embryos after 96-h exposure. The results showed that 5.0 mg/L PCZ induced oxidative damage in zebrafish embryos, as indicated by increased ROS and MDA content and alteration of antioxidative enzyme activity. The activity of Na⁺-K⁺-ATPase in zebrafish embryos was significantly inhibited after exposure to 0.5 mg/L PCZ. The expression levels of bax, p53, casp-3, casp-9, and apaf-1 were significantly increased, indicating that cell apoptosis was caused in embryos by 5.0 mg/L PCZ. The expression level of interleukin-1b (IL-1b) and IL-8 increased after exposure to 0.5 mg/L PCZ, but that of IL-1b, IL-8, and cxcl-c1c (chemokine (C-X-C motif) ligand 18b) decreased in 5.0-mg/L PCZ treatment group, indicating an immunotoxicity effect. Our results suggest that oxidative damage, cell apoptosis, and immunotoxicity would be induced in zebrafish embryos after short-term exposure to PCZ.

The main objective of this study was the development of simple multi-parameter methods for the analyses of biocides in various environmental matrices (wastewater, surface water, and sewage sludge) for measurement and monitoring activities. Eight target substances (triclosan, methyltriclosan (transformation product of triclosan), cybutryne (Irgarol), and the azole fungicides propiconazole, tebuconazole, imazalil, thiabendazole, and cyproconazole) were chosen for determination in selected sample sets. For surface water and wastewater samples a solid-phase extraction (SPE) method and for sewage sludge samples an accelerated solvent extraction (ASE) were developed. The extracts were analyzed by gas chromatographic-mass spectrometric methods (GC/MS), and the analytical methods were checked to ensure sufficient sensitivity by comparing the limits of quantification (LOQs) to the predicted no effect concentrations (PNECs) of the selected biocides. For quality control, recovery rates were determined. Finally, developed methods were checked and validated by application on sample material from various matrices. Sampling took place in seven urban wastewater treatment plants and their corresponding receiving waters. The results revealed that the developed extraction methods are effective and simple and allow the determination of a broad range of biocides in various environmental compartments.

The saprotrophic fungus Alternaria alternata is widespread in the agro-environment and produces more than ten allergenic proteins, mostly protein Alt a 1. The frequency of the Alt a 1 gene was analyzed in a group of A. alternata isolates from winter wheat kernels obtained in Poland, and the effectiveness of various fungicides targeting the pathogen was evaluated. The Alt a 1 gene was identified in four of the seven tested isolates. A. alternata colonized 35.67% kernels on average, but its frequency increased in stored grain where the presence of epiphytes was noted on 23.09 to 51.38% kernels, and endophytes—in 26.21 to 42.01% of kernels. The efficacy of field-applied fungicides did not exceed 50%, despite the fact that A. alternata is highly sensitive to propiconazole, fenpropimorph, and tebuconazole under in vitro conditions. The analyzed isolates were characterized by limited sensitivity to azoxystrobin (EC₅₀ ranged from 0.505 to 1.350 μg cm⁻³) due to a mutation at codon 143 of the CYT b gene, responsible for resistance to quinone outside inhibitor fungicides, which was noted in all isolates. The spread of A. alternata can be effectively controlled with suitable fungicides and by monitoring the prevalence of pathogenic isolates in the environment.