The widespread use of herbicides has increased concern about the hazards and risks to animals living in terrestrial and aquatic ecosystems. A comprehensive understanding of their effective action at different levels of biological organization is critical for establishing guidelines to protect ecosystems and human health. Dinitroanilines are broad-spectrum pre-emergence herbicides currently used for weed control in the conventional agriculture. They are considered extremely safe agrochemicals because they act specifically on tubulin proteins and inhibit shoot and root growth of plants. However, there is a lack of toxicity information regarding the potential risk of exposure to non-target organisms. The aim of the present review is to focus on side effects of the most commonly used active ingredients, e.g. pendimethalin, oryzalin, trifluralin and benfluralin, on animal non-target cells of invertebrates and vertebrates. Acute toxicity varies from slightly to high in terrestrial and aquatic species (i.e. nematodes, earthworms, snails, insects, crustaceans, fish and mammals) depending on the species-specific ability of tested organisms to adsorb and discharge toxicants. Cytotoxicity, genotoxicity and activation of oxidative stress pathways as well as alterations of physiological, metabolic, morphological, developmental and behavioural traits, reviewed here, indicate that exposure to sublethal concentrations of active ingredients poses a clear hazard to animals and humans. Further research is required to evaluate the molecular mechanisms of action of these herbicides in the animal cell and on biological functions at multiple levels, from organisms to communities, including the effects of commercial formulations.

The aim of this study was to monitor the sediment, soil and surface water contamination with selected popular triketone herbicides (mesotrione (MES) and sulcotrione(SUL)), atrazine (ATR) classified as a possible carcinogen and endocrine disrupting chemical, as well as their degradation products, in Silesia (Poland). Seventeen sediment samples, 24 soil samples, and 64 surface water samples collected in 2014 were studied. After solid-liquid extraction (SLE) and solid phase extraction (SPE), analytes were determined by high-performance liquid chromatography (HPLC) with diode array detection (DAD). Ten years after the withdrawal from the use, ATR was not detected in any of the collected samples; however, its degradation products are still present in 41 % of sediment, 71 % of soil, and 8 % of surface water samples. SUL was determined in 85 % of soil samples; its degradation product (2-chloro-4-(methylosulfonyl) benzoic acid (CMBA)) was present in 43 % of soil samples. In 17 % of sediment samples, CMBA was detected. Triketones were detected occasionally in surface water samples. The chemometric analysis (clustering analysis (CA), single-factor analysis of variance (ANOVA), N-Way ANOVA) was applied to find relations between selected soil and sediment parameters and herbicides concentration. In neither of the studied cases a statistically significant relationship between the concentrations of examined herbicides, their degradation products and soil parameters (organic carbon (OC), pH) was observed.

The herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) is widely used in agriculture to control various weeds. The objective of this study was to use the digital image processing method to identify alveolar lesions in the lungs of rats submitted to chronic 2,4-dichlorophenoxyacetic acid (2,4-D) inhalation exposure. We used forty adult male Wistar rats. The rats were divided into four groups: control group (CG), low concentration group (LCG), medium concentration group (MCG), and high concentration group (HCG). In a 6-month exposure period, we used two boxes connected to ultrasonic nebulizers for herbicide spraying. After this period, the rats were euthanized for the collection and study of lung tissue. For each image, counts of injuries and blisters were performed automatically using a methodology based on digital image processing techniques. For analysis of the results, an electronic database (Excel®) was created. We used the Pearson method for correlation analysis; values of p <0.05 were considered significant. In the evaluation of healthy alveoli, we recorded positive and significant correlations between analysis from a pathologist and computational analysis. In the evaluation of injured alveoli, we recorded a positive but non-significant correlation between analysis from a pathologist and computational analysis. These results show the effectiveness of digital image processing when evaluating alveolar integrity.

Acidic herbicides are used to control broad-leaved weeds. They are stable, water-soluble, and with low binding to soil are found frequently in surface waters, often at concentrations above the EU Drinking Water Directive limit of 0.10 μg L-1. This presents a problem when such waters are abstracted for potable supplies. Understanding their sources, transport and fate in river catchments is important. We developed a new Chemcatcher® passive sampler, comprising a 3M Empore™ anion-exchange disk overlaid with a polyethersulphone membrane, for monitoring acidic herbicides (2,4-D, dicamba, dichlorprop, fluroxypyr, MCPA, MCPB, mecoprop, tricolpyr). Sampler uptake rates (Rs = 0.044-0.113 L day-1) were measured in the laboratory. Two field trials using the Chemcatcher® were undertaken in the River Exe catchment, UK. Time-weighted average (TWA) concentrations of the herbicides obtained using the Chemcatcher® were compared with concentrations measured in spot samples of water. The two techniques gave complimentary monitoring data, with the samplers being able to measure stochastic inputs of MCPA and mecoprop occurring in field trial 1. Chemcatcher® detected a large input of MCPA not found by spot sampling during field trial 2. Devices also detected other pesticides and pharmaceuticals with acidic properties. Information obtained using the Chemcatcher® can be used to develop improved risk assessments and catchment management plans and to assess the effectiveness of any mitigation and remediation strategies.

In Colombia, glyphosate (GP) is used to control weeds, with Roundup Active® being the most widely used. This use has affected aquatic ecosystems, causing malformations in amphibians. The Savannah frog (D. molitor) is a tropical frog inhabiting the mountain of Colombia. In the present study, we determined the effect of sublethal concentrations of GP (Roundup Active®) on the skin of D. molitor. Twenty-four tadpoles were exposed to concentrations of GP (T1: 0, T2: 1.4, T3: 3.6, and T4: 5.6 a.e mg/L) during 31 days. In 10 individuals per treatment, two skin regions were evaluated: dorsal cranial and caudal ventral to determine histopathological alterations. Morphometric analysis of the layers of the skin was performed: epidermis, dermis, and hypodermis-muscular. T1 did not present histopathological alterations. Since T2 was identified, glandular cell hyperplasia and hypertrophy increased melanophores and melanin accumulations in the highest concentrations of GP. The ultrastructure revealed an increase in excretory glands in the dermis. In the other layers, an increase of melanophores and melanocyte clusters was observed accompanied by vacuolization of basal cells. The morphometry showed an increase in the thickness of the dermis in the dorso-cranial region in T2 compared to the other treatments, while the ventral caudal region exhibited a variation in the thickness of the dermis from T2 and a decrease in T4. Despite evaluating sublethal concentrations, the skin of D. molitor tadpoles presents histopathological, ultrastructural, and morphometric alterations that could affect the survival of the species in the natural environment.

Herbicides are increasingly applied in vineyards worldwide. However, not much is known on potential side effects on soil organisms or on the nutrition of grapevines (Vitis vinifera). In an experimental vineyard in Austria, we examined the impacts of three within-row herbicide treatments (active ingredients: flazasulfuron, glufosinate, glyphosate) and mechanical weeding on grapevine root mycorrhization; soil microorganisms; earthworms; and nutrient concentration in grapevine roots, leaves, xylem sap and grape juice. The three herbicides reduced grapevine root mycorrhization on average by 53% compared to mechanical weeding. Soil microorganisms (total colony-forming units, CFU) were significantly affected by herbicides with highest CFUs under glufosinate and lowest under glyphosate. Earthworms (surface casting activity, density, biomass, reproduction) or litter decomposition in soil were unaffected by herbicides. Herbicides altered nutrient composition in grapevine roots, leaves, grape juice and xylem sap that was collected 11 months after herbicide application. Xylem sap under herbicide treatments also contained on average 70% more bacteria than under mechanical weeding; however, due to high variability, this was not statistically significant. We conclude that interdisciplinary approaches should receive more attention when assessing ecological effects of herbicides in vineyard ecosystems.

From 2001 to 2014, 13 surveys were conducted in the Baltic Sea, to determine its pollution of 50 micropollutants. The investigations focused mostly on the German western Baltic Sea; in 2008, one survey covered the entire Baltic Sea. Various groups of herbicides (such as triazines, phenoxyacetic acid, phenylurea), perfluoroalkyl substances, pharmaceuticals, and industrial products were analyzed during these surveys. The highest concentrations (median 1 to 4 ng/L) were observed for atrazine, simazine, chloridazone, 2,4-dichlorophenoxyacetic acid, benzotriazole, primidone, and carbamazepine. Most micropollutants exhibited a relatively homogenous spatial distribution, though some herbicides show elevated concentrations in certain regions (e.g., Odra estuary), indicating a riverine input. The data set was analyzed, both for seasonal influences and long-time trends. Some herbicides exhibited higher concentrations during summertime. Both upward- and downward-directed time trends could be identified for some herbicides and perfluorinated compounds. For most of the detected compounds, a low-risk quotient was calculated. Only the occurrence of carbendazim could potentially pose a higher risk to the Baltic Sea.

Simple and effective extraction methods based on matrix solid-phase dispersion (MSPD), dispersive liquid-liquid microextraction (DLLME), and solid-phase extraction (SPE) coupled with high-performance liquid chromatography with diode array detector (HPLC-DAD) were developed to determine triketone herbicides-sulcotrione (SUL), mesotrione (MES), tembotrione (TEMB), and their degradation products-in plant tissues and water samples. The extraction procedures were employed to enable quantification of the accumulation of selected triketone herbicides and their degradation products in a model aquatic plant, Egeria densa. To obtain comprehensive information about the triketones' influence on an aquatic plant, changes in chlorophyll concentration in plants exposed to these triketones were monitored. The average recovery ranged from 58 to 115 % (coefficients of variation 7-12 %) for plant tissues and from 52 to 96 % (coefficients of variation 8-20 %) for water samples. The limit of detection (LOD) for the MSPD-HPLC-DAD procedure was in the range of 0.06-0.23 μg/g, whereas for DLLME-HPLC-DAD and SPE-HPLC-DAD, LOD was in the range of 0.06-0.26 μg/mL. Symptoms of the phytotoxicity of sulcotrione, mesotrione, tembotrione, and their degradation products (decrease of chlorophyll concentration in plant sprouts) were observed for E. densa cultivated in water with herbicide concentrations of 100 μg/L. Moreover, the tembotrione degradation product exhibited a high level of accumulation and low metabolism in plant tissues in comparison to the other triketones and their degradation products.

Pesticide contamination of bee products is a widespread phenomenon. Due to its composition, bee bread is affected by both lipophilic and hydrophilic substances. As proof of concept of a monitoring campaign and to better understand the extent of contamination, we developed an analytical method based on a modified QuEChERS extraction, with subsequent separation by liquid chromatography and detection by mass spectrometry. This allowed for the quantitation of 51 agricultural- or beekeeping-associated pesticides in bee bread. The workflow was applied to 60 samples taken biweekly throughout spring to autumn 2022 from five colonies at a Swiss apiary in an agricultural area. In total, 30 pesticides were identified (> LOD), among which 26 pesticides were quantitated. The total number of pesticides detected per colony ranged from 11 to 19. The most prevalent substances (> LOQ) were two neonicotinoid insecticides, acetamiprid and thiacloprid (max. 16 μg/kg and 37 μg/kg, respectively); seven fungicides, azoxystrobin (max. 72 μg/kg), boscalid (max. 50 μg/kg), cyprodinil (max. 1965 μg/kg), difenoconazole (max. 73 μg/kg), mandipropamid (max. 33 μg/kg), pyraclostrobin (max. 8 μg/kg) and trifloxystrobin (max. 38 μg/kg); and two herbicides, prosulfocarb (max. 38 μg/kg) and terbuthylazine (max. 26 μg/kg). The study revealed strong variability in pesticide occurrence and concentrations among colonies sampled at the same site and date. The applied biweekly sampling of bee bread from March to August was shown to be reliable in capturing peak contaminations and revealing the onset of certain pesticides in bee bread. The study provides an adequate practical approach for pesticide monitoring campaigns.

The combined effects of the herbicide glyphosate and elevated temperature were studied on the tropical staghorn coral Acropora formosa, in Nha Trang bay, Vietnam. The corals were collected from two different reefs, one close to a polluted fish farm and one in a marine-protected area (MPA). In the laboratory, branches of the corals were exposed to the herbicide glyphosate at ambient (28 °C) and at 3 °C elevated water temperatures (31 °C). Effects of herbicide and elevated temperature were studied on coral bleaching using photography and digital image analysis (new colorimetric method developed here based on grayscale), chlorophyll a analysis, and symbiotic dinoflagellate (Symbiodinium, referred to as zooxanthellae) counts. All corals from the MPA started to bleach in the laboratory before they were exposed to the treatments, indicating that they were very sensitive, as opposed to the corals collected from the more polluted site, which were more tolerant and showed no bleaching response to temperature increase or herbicide alone. However, the combined exposure to the stressors resulted in significant loss of color, proportional to loss in chlorophyll a and zooxanthellae. The difference in sensitivity of the corals collected from the polluted site versus the MPA site could be explained by different symbiont types: the resilient type C3u and the stress-sensitive types C21 and C23, respectively. The additive effect of elevated temperatures and herbicides adds further weight to the notion that the bleaching of coral reefs is accelerated in the presence of multiple stressors. These results suggest that the corals in Nha Trang bay have adapted to the ongoing pollution to become more tolerant to anthropogenic stressors, and that multiple stressors hamper this resilience. The loss of color and decrease of chlorophyll a suggest that bleaching is related to concentration of chloro-pigments. The colorimetric method could be further fine-tuned and used as a precise, non-intrusive tool for monitoring coral bleaching in situ.