Insights into interaction of triazine herbicides with three kinds of different alkyl groups (simetryne, ametryn and terbutryn) with human serum albumin via multi-spectral analysis.

In this study, the interaction of triazine herbicides with three kinds of different alkyl groups (simetryne, ametryn and terbutryn) with human serum albumin (HSA) are investigated through UV-vis, fluorescence, and circular dichroism (CD) spectra. The mechanisms on the fluorescence quenching of HSA initiated by triazine herbicides are obtained using Stern-Volmer, Lineweaver-Burk and Double logarithm equations. The quenching rate constant (Kq), Stern-Volmer quenching constant (Ksv), binding constant (KA), thermodynamic parameters such as enthalpy change (∆H), entropy change (∆S) and Gibbs free energy (∆G) and number of binding site (n) are calculated and compared. The variations in the microenvironment of amino acid residues are studied by synchronous fluorescence spectroscopy. The binding sites and subdomains are identified using warfarin and ibuprofen as site probes. The conformational changes of HSA are measured using CD spectra. The results reveal that the triazine herbicides with different alkyl groups can interact with HSA by static quenching. The combination of the three herbicides and HSA are equally proportional, and the binding processes are spontaneous. Hydrophobic interaction forces play important roles in simetryne-HSA and ametryn-HSA, while the interaction of terbutryn-HSA is Van der Waals forces and hydrogen bonding. Moreover, the three herbicides can bind to HSA at site I (sub-domain IIA) more than site II (subdomain IIIA), and combine with tryptophan (Trp) more easily than tyrosine (Tyr) residues, respectively. By comparison, the order of interaction strength is terbutryn-HSA > ametryn-HSA > simetryne-HSA. Terbutryn can destroy the secondary structure of HSA more than simetryne and ametryn, and the potential toxicity of terbutryn is higher. It is expected that the interactions of triazine herbicides with HSA via multi-spectral analysis can offer some valuable information for studying the toxicity and the harm of triazine herbicides on human health at molecular level in life science.

Impact of combined seawater warming and triazine-type herbicide pollution on the physiology and potential toxicity of the dinoflagellate Alexandrium minutum.

Coastal phytoplankton communities are often exposed to multiple anthropogenic stressors simultaneously. Here, we experimentally examined how temperature increase (20-26 °C) and triazine-type herbicides pollution (500 ng terbutryn L-1), both recognized as emerging stressors, affect the abundance, physiology and selected saxitoxin gene expression in the toxic dinoflagellate Alexandrium minutum. The results show that A. minutum is more susceptible to terbutryn pollution with increasing temperatures, resulting in a significant decline in its abundance (∼80 %) and photosynthetic activity (∼40 %), while saxitoxin gene expression increased (1.5-2.5-fold). This suggests that in warming polluted coastal areas where A. minutum is often found, saxitoxin poisoning may occur even in the absence of a massive bloom. Our results recommend the development of science-based monitoring practices for algal dissolved toxins in coastal waters and estuaries, supporting environmental policies under warming and contaminated coastal regions.

Terbutryn causes developmental toxicity in zebrafish (Danio rerio) via apoptosis and major organ malformation in the early stages of embryogenesis.

Terbutryn (2-(ethylamino)-4-(tert-butylamino)-6-(methylthio)-1,3,5-triazine) is a substituted symmetrical triazine herbicide used in agricultural fields to prevent undesired vegetation growth by inhibiting photosynthesis in target weeds. Although terbutryn has various benefits, long-term exposure, misuse, or abuse of terbutryn may cause non-target toxicity and severe ecosystem pollution. To provide a detailed description of the embryonic developmental toxicity of terbutryn, zebrafish (Danio rerio) were exposed to 2, 4, and 6 mg/L of terbutryn and the morphological changes, pathological abnormalities, and developmental endpoints were assessed relative to that of a solvent control. The results showed that terbutryn induces a loss of survivability, reduction in body and eye size, and edema in the yolk sac. Through fluorescence microscopy, blood vessels, motor neurons, and liver development were investigated using transgenic zebrafish models based on fluorescently tagged genes (fllk1:eGFP, olig2:dsRed, and L-fabp:dsRed). Furthermore, cell death by apoptosis in zebrafish caused by terbutryn exposure was evaluated via acridine orange staining, which is a selective fluorescent staining agent. To support the preceding results, gene expression alterations caused by terbutryn exposure in zebrafish larvae were assessed. The overall results indicate that exposure to terbutryn induces apoptosis and disrupts organ development. These embryonic developmental toxicity results suggest that terbutryn should be applied in the right areas at the appropriate rates, concentrations, and quantities.

Transformation and stable isotope fractionation of the urban biocide terbutryn during biodegradation, photodegradation and abiotic hydrolysis.

Terbutryn is a widely used biocide in construction materials like paint and render to prevent the growth of microorganisms, algae and fungi. Terbutryn is released from the facades into the environment during rainfall, contaminating surface waters, soil and groundwater. Knowledge of terbutryn dissipation from the facades to aquatic ecosystems is scarce. Here, we examined in laboratory microcosms degradation half-lives, formation of transformation products and carbon and nitrogen isotope fractionation during terbutryn direct (UV light with λ = 254 nm and simulated sunlight) and indirect (simulated sunlight with nitrate) photodegradation, abiotic hydrolysis (pH = 1, 7 and 13), and aerobic biodegradation (stormwater pond sediment, soil and activated sludge). Biodegradation half-lives of terbutryn were high (>80 d). Photodegradation under simulated sunlight and hydrolysis at extreme pH values indicated slow degradability and accumulation in the environment. Photodegradation resulted in a variety of transformation products, whereas abiotic hydrolysis lead solely to terbutryn-2-hydroxy in acidic and basic conditions. Biodegradation indicates degradation to terbutryn-2-hydroxy through terbutryn-sulfoxide. Compound-specific isotope analysis (CSIA) of terbutryn holds potential to differentiate degradation pathways. Carbon isotope fractionation values (εC) ranged from -3.4 ± 0.3‰ (hydrolysis pH 1) to +0.8 ± 0.1‰ (photodegradation under UV light), while nitrogen isotope fractionation values ranged from -1.0 ± 0.4‰ (simulated sunlight photodegradation with nitrate) to +3.4 ± 0.2‰ (hydrolysis at pH 1). In contrast, isotope fractionation during biodegradation was insignificant. ΛN/C values ranged from -1.0 ± 0.1 (hydrolysis at pH 1) to 2.8 ± 0.3 (photodegradation under UV light), allowing to differentiate degradation pathways. Combining the formation of transformation products and stable isotope fractionation enabled identifying distinct degradation pathways. Altogether, this study highlights the potential of CSIA to follow terbutryn degradation in situ and differentiate prevailing degradation pathways, which may help to monitor urban biocide remediation and mitigation strategies.

A reliable LC-MS/MS-based method for trace level determination of 50 medium to highly polar pesticide residues in sediments and ecological risk assessment.

The occurrence of polar pesticides in sediments has not been extensively investigated because of their relatively poor hydrophobicity and apparently less persistence in the environment. However, their continuous release into the aquatic systems calls for the evaluation of their potential accumulation in sediments and the role of this matrix as a potential source of these compounds. Considering this, a method based on pressurized liquid extraction (PLE), extract clean-up by solid phase extraction (SPE), and analyte determination by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed and validated to analyze 50 relevant (frequently used and/or regulated or found in water) medium to highly polar pesticides in sediments. The method showed good performance regarding accuracy (relative recoveries between 76 and 124%), precision (relative standard deviation values < 20%), sensitivity (LODs in the low nanogram per gram for most compounds), linearity (coefficients of determination > 0.99), and matrix effects (negligible for all analytes). The use of an isotope dilution approach for quantification ensures result reliability. As a part of the validation process, the method was applied to the analysis of the target pesticides in sediments from the Llobregat River (NE Spain) showing the presence of five of them, namely, terbutryn, dichlorvos, terbuthylazine, diazinon, and irgarol. All 5 pesticides, due to both the concentrations found and their physical-chemical characteristics, demonstrate high potential for bioaccumulation and risk to aquatic organisms. Additional multi-disciplinary studies that investigate pesticide occurrence in different aquatic compartments and evaluate the potential risks for aquatic ecosystems are required to assess the environmental impact and significance of the presence of pesticides in sediments. Graphical Abstract.

Laboratory-to-field extrapolation: Increase in carbamazepine toxicity in a higher tier, multiple-stress experiment.

The toxicity and environmental risk of chemicals, such as the antiepileptic drug carbamazepine (CBZ), is commonly assessed using standardized laboratory tests and laboratory-to-field extrapolation. To investigate the toxicity of CBZ to aquatic key organisms in a more complex and environmentally relevant scenario, we conducted a 32-day multiple-stress experiment in artificial indoor streams. We exposed the non-biting midge Chironomus riparius, the blackworm Lumbriculus variegatus, and the New Zealand mud snail Potamopyrgus antipodarum to 80 and 400 µg CBZ/L in six artificial indoor streams. In addition to hydraulic stress, species' interaction, and low organic content in the sediment, organisms were co-exposed to the herbicide terbutryn (TBY) as a second chemical stressor at a concentration of 6 µg/L. The exposure to CBZ under multiple stress conditions resulted in a 10- to more than 25-fold higher toxicity in C. riparius and P. antipodarum when compared to a previous, standardized laboratory experiment. The co-exposure to TBY enhanced the adverse effects of CBZ on snails (reduced production of embryos). This effect was additive as the single exposure to TBY also reduced the reproduction of snails, most likely through the reduction of biofilm biomass. The emergence of C. riparius declined at a CBZ concentration of 400 µg/L (without the co-exposure to TBY) and at 80 µg/L in combination with TBY. The difference in sensitivity between laboratory and indoor stream experiments is indicative of a potential underestimation of risk when toxicity data are extrapolated to field conditions. The present results suggest the inclusion of non-chemical and chemical stressors in environmental hazard and risk assessments.

Influence of pigments on phototransformation of biocides in paints.

Biocides are commonly applied to construction materials such as facade renders and paints in order to protect them from microbial spoilage. These renders and paints are exposed to weathering conditions, e.g., sunlight and rain. Pigments are interacting intensively with the spectrum of the incoming light; thus, an effect of paint pigments on phototransformation rates and reaction pathways of the biocides is hypothesized. In this study, the phototransformation of four commonly used biocides (carbendazim, diuron, octylisothiazolinone (OIT) and terbutryn) in four different paint formulations differing solely in pigments (red and black iron oxides, white titanium dioxide, and one pigment-free formulation) were investigated. Paints surfaces were irradiated under controlled conditions. The results show that biocides degrade most rapidly in the pigment-free formulation. The degradation in the pigment-free formulation followed a first-order kinetic model with the respective photolysis rate constants: kp,Diuron = 0.0090 h-1, kp,OIT = 0.1205 h-1, kp,Terbutryn = 0.0079 h-1. Carbendazim concentrations did not change significantly. The degradation was considerably lower in the pigment-containing paints. The determination of several phototransformation products of terbutryn and octylisothiazolinone showed different transformation product ratios dependent on the pigment. Consequently, pigments not only reflect the incoming light, but also interact with the biocide photodegradation.

Direct and indirect effects of pesticides on a benthic grazer during its life cycle.

BACKGROUND: Macroinvertebrates in aquatic ecosystems are repeatedly exposed to pesticides during their life cycle. Effects of consecutive exposure during different life stages and possible synergistic effects are not addressed in the standardized hazard assessment. The present study investigated two environmentally relevant exposure scenarios in batch (microcosm) and artificial indoor stream (mesocosm) experiments using the larvae of the mayfly Rhithrogena semicolorata (grazer) and natural aufwuchs. Grazers were analysed regarding growth, physiological condition, and drift behaviour, while the aufwuchs was analysed in terms of biomass using the particulate organic carbon as well as the chlorophyll a content. The aim was to reveal direct and indirect effects of an herbicide exposure during autumn on juvenile grazers and an insecticide exposure during spring on semi-juvenile grazers. RESULTS: Direct and indirect effects were found in both exposure scenarios at environmentally relevant concentrations. In the herbicide exposure scenario with terbutryn, clear direct effects on the aufwuchs community with a LOEC of 0.38 µg L-1 were found. Effect levels of grazers due to indirect effects were equal, with the overnight drift being the most sensitive grazer endpoint. In the insecticide exposure scenario, clear lethal and sub lethal effects of lambda-cyhalothrin were evident. Derived LC50 values for the artificial indoor stream and batch experiment were 2.42 µg g-1 OC (69 days) and 1.2 µg g-1 OC (28 days), respectively. Sub lethal effects in terms of increased drift as well-reduced growth and triglyceride levels were found at concentrations of 1.4 and 0.09 µg g-1 OC (LOECs). These results were confirmed by the batch experiment, which revealed effect values in the similar range. Finally, a clear indirect effect of the insecticide on the aufwuchs was evident in the batch experiment with an LOEC at 0.9 µg g-1 OC. CONCLUSION: Toxicity Exposure Ratios calculated with the derived effect values indicate a risk for the investigated grazer by both pesticides. Moreover, observed indirect effects during the herbicide exposure seem to be able to affect the grazers during a second exposure with an insecticide, due to reduced physiological conditions. We suggest further research with time-shifted exposure scenarios to gain a better understanding of the complex interactions of pesticides with the life cycle and the food webs of macroinvertebrates.

Analytical method validation for terbutryn using gas chromatography/ion trap, gas chromatography/mass selective detector, and liquid chromatography/triple quadrupole mass spectrometers.

Analytical methods including solvent extraction followed by gas chromatography/ion-trap (GC/IT) with scan and MS/MS mode, a GC/mass selective detector (GC/MSD), and liquid chromatography/triple quadrupole mass spectrometers (LC/MS/MS) were optimized to identify and quantify terbutryn. The spike recovery was 96.5% using GC/IT with scan mode and 103.5% with MS/MS mode, 90.3% by GC/MSD, and 92.5% by LC/MS/MS. The limit of detection (LOD) was 0.0015 mg/kg by GC/IT with scan, 0.026 mg/kg with MS/MS mode, 0.015 mg/kg with GC/MSD, and 0.026 mg/kg by LC/MS/MS. Of the four methods, GC/IT with scan mode was determined to be the most sensitive (with LOD: 0.0015 mg/kg and limit of quantitation (LOQ): 0.0047 mg/kg), rapid (retention time: 9.6 min) and the most precise method (relative standard deviation: 17%) for the quantification of terbutryn. GC/IT with scan mode proved to be the more sensitive analytical method for terbutryn than other methods in this study, showing better accuracy and rapid analysis.

Development of a Matrix Solid Phase Dispersion methodology for the determination of triazine herbicides in mussels.

A method based on Matrix Solid Phase Dispersion (MSPD) for determination of nine triazines in mussels has been optimised in terms of the sorbents used for extracting and cleaning-up. Two dispersing agents: C18 and florisil, and eight cleanup co-sorbents: florisil, silica, silica/alumina, Envi™ Carb, Envi-Carb-II/PSA, SAX/PSA, Envi-Carb-II /SAX/PSA and C18 were assayed. Analytes were eluted using 20 mL of ethyl acetate and 5 mL of acetonitrile and finally the extract was concentrated to dryness, re-constituted with 1 mL methanol and determined by HPLC-DAD. The best results were obtained with C18 as dispersing agent and Envi-Carb-II/SAX/PSA as clean-up co-column. Recoveries ranged between 79% and 99% and repeatability and reproducibility were below than 16% for all compounds. The linearity of the calibration curves yielded the R(2)⩾0.9993. The LOQ values ranged from 0.10 to 0.18 mg kg(-1) dried sample. Finally the method was applied to the analysis of mussel samples from Galicia (NW Spain).

Dynamics of biocide emissions from buildings in a suburban stormwater catchment - concentrations, mass loads and emission processes.

Biocides such as isothiazolinones, carbamates, triazines, phenylureas, azoles and others are used to protect the surfaces of buildings, e.g. painted or unpainted render or wood. These biocides can be mobilized from the materials if rainwater gets into contact with these buildings. Hence, these biocides will be found in rainwater runoff (stormwater) from buildings that is traditionally managed as "clean water" in stormwater sewer systems and often directly discharged into surface waters without further treatment. By means of a 9 month event-based high resolution sampling campaign the biocide emissions in a small suburban stormwater catchment were analysed and the emission dynamics throughout the single rain events were investigated. Five out of twelve of the rain events (peak events) proved significantly higher concentrations than the rest (average) for at least one compound. Highest median concentrations of 0.045 and 0.052 µg L(-1) were found for terbutryn and carbendazim, while the concentrations for isoproturon, diuron, N-octylisothiazolinone, benzoisothiazolinone, cybutryn, propiconazole, tebuconazole, and mecoprop were one order of magnitude lower. However, during the peak events the concentrations reached up to 1.8 and 0.3 µg L(-1) for terbutryn and carbendazim, respectively. Emissions of an averaged single family house into the stormwater sewer turned out to be 59 and 50 µg event(-1) house(-1) terbutryn and carbendazim, respectively. Emissions for the other biocides ranged from 0.1 to 11 µg event(-1) house(-1). Mass load analysis revealed that peak events contributed in single events as much to the emissions as 11 average events. However, the mass loads were highly dependent on the amounts of rainwater, i.e. the hydraulic flow in the receiving sewer pipe. The analysis of the emission dynamics showed first flush emissions only for single parameters in three events out of twelve. Generally biocides seemed to be introduced into the stormwater system rather continuously during the respective events than in the beginning of them. Mass flows during the events did correlate to driving rain, whereas mass loads neither correlated to the length or the intensity of rainfall nor the length of dry period.

Removal of triazine herbicides from aqueous systems by a biofilm reactor continuously or intermittently operated.

The impact of pesticide movement via overland flow or tile drainage water on the quality of receiving water bodies has been a serious concern in the last decades; thus, for remediation of water contaminated with herbicides, bioreaction systems designed to retain biomass have been proposed. In this context, the aim of this study was to evaluate the atrazine and terbutryn biodegradation capacity of a microbial consortium, immobilized in a biofilm reactor (PBR), packed with fragments of porous volcanic stone. The microbial consortium, constituted by four predominant bacterial strains, was used to degrade a commercial formulation of atrazine and terbutryn in the biofilm reactor, intermittently or continuously operated at volumetric loading rates ranging from 44 to 306 mg L(-1) d(-1). The complete removal of both herbicides was achieved in both systems; however, higher volumetric removal rates were obtained in the continuous system. It was demonstrated that the adjuvants of the commercial formulation of the herbicide significantly enhanced the removal of atrazine and terbutryn.