Au@PtPd enhanced immunoassay with 3D printed smartphone device for quantification of diaminochlorotriazine (DACT), the major atrazine biomarker.

Increased use of pesticides in agriculture requires new advanced techniques to monitor both environmental levels and human exposure of pesticides to avoid potential adverse health outcomes in sensitive populations. Atrazine is widely used to control broadleaf weeds, and here we developed a new sensor capable of detecting diaminochlorotriazine (DACT), the major metabolite and biomarker of atrazine exposure. We established an Au@PtPd nanoparticles labeled lateral flow immunoassay (LFIA) for immunochromatographic based rapid detection of urinary DACT. The detection was based on competitive immunoassay between the analyte and the BSA-conjugated antigen. As evaluated, the coupled mesoporous core-shell Au@PtPd nanoparticles, with superior peroxidase-like activity, as the signal indicator offers a rapid direct chromatographic readout inversely correlated with the concentration of analytes, providing a detection limit of 0.7 ng/mL for DACT. Moreover, the detection limits were boosted to as low as 11 pg/mL with the detectable range from 10 pg/ml to 10 ng/mL, through a one-step catalytic chromogenic reaction. A rapid readout device was developed by 3D printing to provide a stable real-time quantification of the color intensity capable of assessing both chromatographic and absorbance results. This Au@PtPd nanoparticle-based immunosensing platform, as well as the 3D printed readout device, provide a promising tool for on-site and ultrasensitive detection of pesticide biomarkers.

Causal factors for pesticide trends in streams of the United States: Atrazine and deethylatrazine.

Pesticides are important for agriculture in the United States, and atrazine is one of the most widely used and widely detected pesticides in surface water. A better understanding of the mechanisms by which atrazine and its degradation product, deethylatrazine, increase and decrease in surface waters can help inform future decisions for water quality improvement. This study considers causal factors for trends in pesticide concentration in U.S. streams and models the causal factors, other than use, in structural equation models. The structural equation models use a concomitant trend in corn (Zea mays L.) and a latent variable model, indicating moisture supply and management. The moisture supply and management latent variable model incorporates long-term moisture conditions in the individual watersheds by using the Palmer hydrologic drought index, human influence on the hydrologic cycle through the percentage of the watershed drained by tile drains in 2012, and the base-flow contribution to streamflow, using the base-flow index. The structural equation models explain 77 and 38% of the variability in atrazine and deethylatrazine trends, respectively, across the conterminous United States. The models highlight future water quality challenges, particularly in tile-drained settings where fall precipitation and heavy precipitation are increasing.

A nationwide study of the occurrence and distribution of atrazine and its degradates in tap water and groundwater in China: Assessment of human exposure potential.

Despite frequent detection of atrazine (ATZ) and its degradates (including hydroxyatrazine, ATZ-OH; deethylatrazine, DEA; deisopropylatrazine, DIA; and deethyldeisopropylatrazine, DACT) in a variety of water bodies, documentation of their occurrence and distribution in tap water in China is still scarce. A nationwide survey about ATZ and its degradates (ATZs) in tap water from 31 provinces in 7 regions of mainland China and Hong Kong was conducted during June 2019. At least one of the analytes was found in all the water samples (n = 884). The median sum concentrations of ATZs (ΣATZs) was 21.0 ng/L (range: 0.02 ng/L-3.04 µg/L). The predominant compounds of ATZs in tap water were ATZ and DEA, with a detection frequency of 99.5% and 98.0%, respectively, followed by ATZ-OH (87.3%), DACT (84.0%), and DIA (78.1%). Significant regional variations (p < 0.05) were found in the concentrations of ATZs in tap water, and the highest concentration of ΣATZs (median: 254 ng/L, range: 0.44 ng/L-3.04 µg/L) was found in Northeastern China, followed by Eastern (37.2 ng/L, 0.02-706 ng/L), Northern (30.2 ng/L, 0.04-317 ng/L), Central (29.3 ng/L, 0.04-256 ng/L), Southern (25.0 ng/L, 0.04-297 ng/L), Southwestern (17.2 ng/L, 0.02-388 ng/L), and Northwestern China (3.22 ng/L, 0.06-214 ng/L). The level of ΣATZs in groundwater from rural area of China was about 1/3 of that found in tap water. ATZs cannot be removed by boiling tap water. The highest estimated daily intake of ΣATZs (248 ng/kg-body weight/day) was found in the infant population of Changchun, Jilin, Northeastern China.

The evolving story of AtzT, a periplasmic binding protein.

Atrazine is an s-triazine-based herbicide that is used in many countries around the world in many millions of tons per year. A small number of organisms, such as Pseudomonas sp. strain ADP, have evolved to use this modified s-triazine as a food source, and the various genes required to metabolize atrazine can be found on a single plasmid. The atomic structures of seven of the eight proteins involved in the breakdown of atrazine by Pseudomonas sp. strain ADP have been determined by X-ray crystallography, but the structures of the proteins required by the cell to import atrazine for use as an energy source are still lacking. The structure of AtzT, a periplasmic binding protein that may be involved in the transport of a derivative of atrazine, 2-hydroxyatrazine, into the cell for mineralization, has now been determined. The structure was determined by SAD phasing using an ethylmercury phosphate derivative that diffracted X-rays to beyond 1.9 Šresolution. `Native' (guanine-bound) and 2-hydroxyatrazine-bound structures were also determined to high resolution (1.67 and 1.65 Å, respectively), showing that 2-hydroxyatrazine binds in a similar way to the purportedly native ligand. Structural similarities led to the belief that it may be possible to evolve AtzT from a purine-binding protein to a protein that can bind and detect atrazine in the environment.

Transfer and degradation of the common pesticide atrazine through the unsaturated zone of the Chalk aquifer (Northern France).

Groundwater in the Chalk aquifer is an important water resource whose quality has degraded due to fertilizer and pesticide use. Atrazine, classified as a priority substance, has been one of the most applied pesticides and also one of the most frequently detected pesticides in groundwater. The present study investigated the transfer and degradation of atrazine in the unsaturated zone of the Chalk aquifer in Northern France. The study was conducted in an underground quarry (Saint-Martin-le-Noeud), which provides a direct access to the water table and intercepts the unsaturated zone at different depths. The lake and the ceiling percolation of 16 sites throughout the quarry were followed. For 16 sites, the percolating flow rate and lake level were measured and the lake water was sampled for nitrate, atrazine and deethylatrazine (DEA, main degradation product of atrazine) analysis over 2.5 years. High spatial variations in hydrodynamics (percolating flow rate and lake level) and in lake water quality (atrazine between 55±11 and 202±40 ng L-1 and DEA between 269±53 and 1727±345 ng L-1) indicate that the properties of the unsaturated zone influence the transfer and the degradation of atrazine. A counterclockwise hysteresis characterizes the relationship between the lake level and atrazine concentration. Temporal variation shows that the atrazine is transferred through the matrix and fractures with a delay caused by the sorption process that differs in atrazine and DEA. The layer of clay-with-flints is shown to favor the degradation of atrazine near the surface. Preferential pathways may be created below clay-with-flints, through which the transfer of atrazine is quicker.

Effect of the herbicide atrazine and its major metabolite, DACT, on bovine sperm cryotolerance.

During freezing and thawing procedures, sperm are exposed to chemical and/or physical stressors that may cause adverse and harmful changes to sperm membranes. Accurate evaluation of the structural and functional integrity of fresh as well as cryopreserved sperm is highly important in predicting sperm fertilization capacity and success of artificial insemination (AI). The herbicide atrazine (ATZ) and its major metabolite, diaminochlorotriazine (DACT) are considered a ubiquitous environmental contaminants and endocrine disruptors, which deleteriously effect sperm function. Taking into consideration possible damage caused by environmental contaminants to sperm membranes, additive effects during cryopreservation cannot be ruled out. The aim of the current study was to evaluate the effect of ATZ (0.1 or 1 µM) and DACT (1 or 10 µM) exposure during or after cryopreservation on bovine sperm cryotolerance. Sperm membrane integrity and functionality were evaluated using fluorimetric probes: (1) double-stranded DNA was examined by 4',6-diamidino-2-phenylindole; (2) plasma membrane integrity was examined by propidium iodide; (3) acrosome reaction (AR) was examined by fluorescein isothiocyanate-conjugated Pisum sativum agglutinin; mitochondrial membrane potential (ΔΨm) was examined by 5,5',6,6'-tetra-chloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide fluorescent probe. The findings demonstrate, that exposure of sperm to ATZ (0.1 or 1 µM) or DACT (1 or 10 µM) during cryopreservation increased the proportion of dead sperm relative to the control (P < 0.09); exposure to DACT (1 or 10 µM) increased ΔΨm (P < 0.03). Neither ATZ nor DACT affected spontaneous AR. In contrast, the proportion of sperm with Ca++ ionophore-induced AR was lower after exposure to 1 µM DACT (P < 0.05). Following freezing and thawing procedures, exposing sperm to 1 µM ATZ increased the proportion of dead sperm relative to the control (P < 0.05), but had no significant effect on sperm ΔΨm or AR. In conclusion, exposing sperm to endocrine-disrupting chemicals such as ATZ or DACT during cryopreservation reduces sperm cryotolerance and resistance post-thawing.

Characterization of Activation of the Hypothalamic-Pituitary-Adrenal Axis by the Herbicide Atrazine in the Female Rat.

Atrazine (ATR) is a commonly used pre-emergence and early postemergence herbicide. Rats gavaged with ATR and its chlorometabolites desethylatrazine (DEA) and deisopropylatrazine (DIA) respond with a rapid and dose-dependent rise in plasma corticosterone, whereas the major chlorometabolite, diaminochlorotriazine (DACT), has little or no effect on corticosterone levels. In this study, we investigated the possible sites of ATR activation of the hypothalamic-pituitary-adrenal (HPA) axis. ATR treatment had no effect on adrenal weights but altered adrenal morphology. Hypophysectomized rats or rats under dexamethasone suppression did not respond to ATR treatment, suggesting that ATR does not directly stimulate the adrenal gland to induce corticosterone synthesis. Immortalized mouse corticotrophs (AtT-20) and primary rat pituitary cultures were treated with ATR, DEA, DIA, or DACT. None of the compounds induced an increase in ACTH secretion or potentiated ACTH release in conjunction with CRH on ACTH release. In female rats gavaged with ATR, pretreatment with the CRH receptor antagonist astressin completely blocked the ATR-induced rise in corticosterone concentrations, implicating CRH release in ATR-induced HPA activation. Intracerebroventricular infusion of ATR, DEA, and DIA but not DACT at concentrations equivalent to peak plasma concentrations after gavage dosing resulted in an elevation of plasma corticosterone concentrations. However, ATR did not induce c-Fos immunoreactivity in the paraventricular nucleus of the hypothalamus. These results indicate that ATR activates the HPA axis centrally and requires CRH receptor activation, but it does not stimulate cellular pathways associated with CRH neuronal excitation.

Concentration/time-dependent dissipation, partitioning and plant accumulation of hazardous current-used pesticides and 2-hydroxyatrazine in sand and soil.

The dissipation, partitioning dynamics and biouptake was measured for selected hazardous current-used pesticides (conazole fungicides: epoxiconazole, flusilazole, tebuconazole; prochloraz, chlorpyrifos, pendimethalin) and for a transformation product (2-hydroxyatrazine) in agricultural soil and quartz sand as representatives of a real and a worst-case scenario. Dissipation, uptake to Lactuca sativa and the freely dissolved concentration along with the organic carbon-normalized sorption coefficients (Koc) were determined on days 12, 40, and 90 following the application of compounds at three fortification levels (0.1-1.0-10 mg/kg). Conazole fungicides showed similar dissipation patterns and were more persistent in soil than prochloraz, chlorpyrifos and pendimethalin. 2-Hydroxyatrazine showed a concentration-depended decrease in persistency in soil. Lettuce roots were shown to accumulate higher amounts than shoots where the extent of root uptake was driven by compound partitioning. This was evidenced by the ability of freely dissolved concentration (Cfree) to reliably (r2 = 0.94) predict root uptake. Concentration in leaves did not exceed the maximum residue levels (MRLs) for lettuce, which was likely given by the low root-to-shoot translocation factors (TFs) of the tested compounds varying between 0.007 and 0.14. Koc values were in the range of literature values. Sorption to soil was higher than to sand for all compounds, yet following the Koc dynamics compounds did not appear to be sequestered in soil with increasing residence time. From these results, it follows that the tested compounds may persist in soil but since they did not accumulate in lettuce above MRLs, contamination of the food web is unlikely.

Degradation of atrazine by electrochemically activated persulfate using BDD anode: Role of radicals and influencing factors.

A novel advanced oxidation process using boron-doped diamond (BDD) anode to activate persulfate (PS) with low concentration of electrolyte was systematically investigated in this study. Compared to direct electrochemical oxidation of atrazine (ATZ) using BDD anode, the addition and activation of PS significantly declined the demand for electrolytes. It was confirmed by scavenger experiments that both radical and non-radical oxidation occurred in this system. Degradation of ATZ was enhanced with the increase of current density and dosage of PS, and decrease of initial pH. However, the increase of current density can also lead to the decrease of current efficiency, then increase of energy consumption. Besides, the inhibitory effect of anions on the degradation of ATZ followed the order of HCO3->H2PO4->NO3-, while the presence of Cl- accelerated the degradation of ATZ. Furthermore, the degradation products mainly resulting from de-alkylation, de-chlorination, and hydroxylation were detected. Due to the distinctive preference to ethyl group in BDD/PS system, the formation of deethyl-atrazine was quicker than that of deisopropyl-atrazine. The study aims to provide a comprehensive understanding on the potential application of BDD/PS system in water treatment.

Capillary moving-boundary isotachophoresis with electrospray ionization mass-spectrometric detection and hydrogen ion used as essential terminator: Methodology for sensitive analysis of hydroxyderivatives of s-triazine herbicides in waters.

Capillary isotachophoresis (ITP) is an electrophoretic technique offering high sensitivity due to permanent stacking of the migrating analytes. Its combination with electrospray-ionization mass-spectrometric (ESI-MS) detection is limited by the narrow spectrum of ESI-compatible components but can be compensated by experienced system architecture. This work describes a methodology for sensitive analysis of hydroxyderivatives of s-triazine herbicides, based on implementation of the concepts of moving-boundary isotachophoresis and of H+ as essential terminating component into cationic ITP with ESI-MS detection. Theoretical description of such kind of system is given and equations for zone-related boundary mobilities are derived, resulting in a much more general definition of the effective mobility of the terminating H+ zone than used so far. Explicit equations allowing direct calculation for selected simple systems are derived. The presented theory allows prediction of stacking properties of particular systems and easy selection of suitable electrolyte setups. A simple ESI-compatible system composed of acetic acid and ammonium with H+ and ammonium as a mixed terminator was selected for the analysis of 2-hydroxyatrazine and 2-hydroxyterbutylazine, degradation products of s-triazine herbicides. The proposed method was tested with direct injection without any sample pretreatment and provided excellent linearity and high sensitivity with limits of detection below 100ng/L (0.5nM). Example analyses of unspiked and spiked drinking and river water are shown.

Assessing the phytoremediation potential of crop and grass plants for atrazine-spiked soils.

Pollution of soil and groundwater by atrazine has become an increasing environmental concern in the last decade. A phytoremediation test using plastic pots was conducted in order to assess the ability of several crops and grasses to remove atrazine from a soil of low permeability spiked with this herbicide. Four plant species were assessed for their ability to degrade or accumulate atrazine from soils: two grasses, i.e., ryegrass (Lolium perenne) and tall fescue (Festuca arundinacea), and two crops, i.e., barley (Hordeum vulgare) and maize (Zea mays). Three different doses of atrazine were used for the contamination of the pots: 2, 5 and 10 mg kg-1. 16 days after spiking, the initial amount of atrazine was reduced by 88.6-99.6% in planted pots, while a decrease of only 63.1-78.2% was found for the unplanted pots, thus showing the contribution of plants to soil decontamination. All the plant species were capable of accumulating atrazine and its N-dealkylated metabolites, i.e., deethylatrazine and deisopropylatrazine, in their tissues. Some toxic responses, such as biomass decreases and/or chlorosis, were observed in plants to a greater or lesser extent for initial soil doses of atrazine above 2 mg kg-1. Maize was the plant species with the highest ability to accumulate atrazine derivatives, reaching up to 38.4% of the initial atrazine added to the soil. Rhizosphere degradation/mineralization by microorganisms or plant enzymes, together with degradation inside the plants, have been proposed as the mechanisms that contributed to a higher extent than plant accumulation to explain the removal of atrazine from soils.

Factors controlling spatial and temporal patterns of multiple pesticide compounds in groundwater (Hesbaye chalk aquifer, Belgium).

Factors governing spatial and temporal patterns of pesticide compounds (pesticides and metabolites) concentrations in chalk aquifers remain unclear due to complex flow processes and multiple sources. To uncover which factors govern pesticide compound concentrations in a chalk aquifer, we develop a methodology based on time series analyses, uni- and multivariate statistics accounting for concentrations below detection limits. The methodology is applied to long records (1996-2013) of a restricted compound (bentazone), three banned compounds (atrazine, diuron and simazine) and two metabolites (deethylatrazine (DEA) and 2,6-dichlorobenzamide (BAM)) sampled in the Hesbaye chalk aquifer in Belgium. In the confined area, all compounds had non-detects fractions >80%. By contrast, maximum concentrations exceeded EU's drinking-water standard (100 ng L-1) in the unconfined area. This contrast confirms that recent recharge and polluted water did not reach the confined area, yet. Multivariate analyses based on variables representative of the hydrogeological setting revealed higher diuron and simazine concentrations in the southeast of the unconfined area, where urban activities dominate land use and where the aquifer lacks protection from a less permeable layer of hardened chalk. At individual sites, positive correlations (up to τ=0.48 for bentazone) between pesticide compound concentrations and multi-annual groundwater level fluctuations confirm occurrences of remobilization. A downward temporal trend of atrazine concentrations likely reflects decreasing use of this compound over the last 28 years. However, the lack of a break in concentrations time series and maximum concentrations of atrazine, simazine, DEA and BAM exceeding EU's standard post-ban years provide evidence of persistence. Contrasting upward trends in bentazone concentrations show that a time lag is required for restriction measures to be efficient. These results shed light on factors governing pesticide compound concentrations in chalk aquifers. The developed methodology is not restricted to chalk aquifers, it could be transposed to study other pollutants with concentrations below detection limits.

Immunotoxic effects of atrazine and its main metabolites at environmental relevant concentrations on larval zebrafish (Danio rerio).

Atrazine (ATZ) and its main metabolites, i.e., diaminochlorotriazine (DACT), deisopropylatrazine (DIP), and deethylatrazine (DE), have been widely detected in surface water around the world. In the present study, to determine their immunotoxic effects, zebrafish during the early developmental stage were exposed to ATZ and its main metabolites at environmental concentrations (30, 100, 300 µg L-1). It was observed that ATZ, DACT, DIP and DE selectively induced the transcription of immunotoxic related genes including Tnfα, Il-1ß, Il-6, Il-8, Cxcl-clc and Cc-chem in larval zebrafish. Pretreatment with ATZ and its metabolites also changed the immune response of larval zebrafish to LPS and E. coli challenge, which was indicated by the alternation in the mRNA levels of some cytokines. In addition, 300 µg L-1 ATZ and DACT exposure could also increase the release of tryptase into water, indicating that they increased the anaphylactoid reaction in the larval zebrafish. According to these results, both of ATZ and its metabolites exposure could cause the immunotoxicity in larval zebrafish. Thus, we thought that the ecological risks of the metabolites of ATZ on aquatic organisms could not be ignored.

Spatial distribution of triazine residues in a shallow alluvial aquifer linked to groundwater residence time.

At present, some triazine herbicides occurrence in European groundwater, 13 years after their use ban in the European Union, remains of great concern and raises the question of their persistence in groundwater systems due to several factors such as storage and remobilization from soil and unsaturated zone, limited or absence of degradation, sorption in saturated zones, or to continuing illegal applications. In order to address this problem and to determine triazine distribution in the saturated zone, their occurrence is investigated in the light of the aquifer hydrodynamic on the basis of a geochemical approach using groundwater dating tracers (3H/3He). In this study, atrazine, simazine, terbuthylazine, deethylatrazine, deisopropylatrazine, and deethylterbuthylazine are measured in 66 samples collected between 2011 and 2013 from 21 sampling points, on the Vistrenque shallow alluvial aquifer (southern France), covered by a major agricultural land use. The frequencies of quantification range from 100 to 56 % for simazine and atrazine, respectively (LQ = 1 ng L-1). Total triazine concentrations vary between 15 and 350 ng L-1 and show three different patterns with depth below the water table: (1) low concentrations independent of depth but related to water origin, (2) an increase in concentrations with depth in the aquifer related to groundwater residence time and triazine use prior to their ban, and (3) relatively high concentrations at low depths in the saturated zone more likely related to a slow desorption of these compounds from the soil and unsaturated zone. The triazine attenuation rate varies between 0.3 for waters influenced by surface water infiltration and 4.8 for water showing longer residence times in the aquifer, suggesting an increase in these rates with water residence time in the saturated zone. Increasing triazine concentrations with depth is consistent with a significant decrease in the use of these pesticides for the last 10 years on this area and highlights the efficiency of their ban.

Effect of the herbicide atrazine and its metabolite DACT on bovine sperm quality.

Atrazine (ATZ), one of the most extensively used herbicides, is considered a ubiquitous environmental contaminant. ATZ is a known endocrine disruptor, and deleterious effects on reproductive function have been shown, even at low, ecologically relevant doses (0.1-3µg/L). Once it enters the body, ATZ is metabolized to various metabolites, which are further detected in the urine, serum and tissues. In mammals, the major ATZ metabolite is diaminochlorotriazine (DACT). The current study focuses on direct effects of low doses of ATZ and DACT on bovine sperm isolated from ejaculates or epididymis compartments (head, body and tail). Sperm were incubated under capacitation conditions with or without 0.1-10µM ATZ or 1-100µM DACT. The integrity and functionality of sperm membranes (plasma, acrosomal and mitochondrial) were examined simultaneously by fluorescence staining at 0, 2 and 4h of incubation. Acrosome reaction (AR) was induced by Ca++ ionophore, after capacitation. The findings indicated that both ATZ and DACT adversely affect sperm, expressed by damaged sperm membranes. ATZ had a prominent effect on epididymal-tail sperm, expressed as disruption of all examined membranes, mostly at low (0.1 or 1µM) concentrations; pseudo-AR and that induced by Ca++ ionophore were both affected by exposure to 0.1µM ATZ (P<0.05 and P<0.00004, respectively). A similar pattern was documented for sperm isolated from ejaculates (P<0.002 and P<0.001, respectively). ΔYm was affected by ATZ in sperm isolated from the epididymis tail (1µM, P<0.0009), but not in that isolated from ejaculates. DACT reduced sperm viability at all examined concentrations and in all fractions. DACT at 1µM impaired ΔΨm in sperm isolated from the epididymis tail and ejaculate (P<0.005). DACT at 100µM did not induce pseudo-AR in sperm isolated from the ejaculate, but did in sperm isolated from the epididymis tail (P<0.05). Induction of AR by Ca++ ionophore was impaired in sperm isolated from ejaculate and exposed to 10 or 100µM DACT (P<0.05) and in sperm isolated from the epididymis tail and exposed to 1, 10 or 100µM DACT (P<0.0004). These findings reveal the harmful effect of exposure to ATZ and DACT, mainly at low ecologically relevant doses, on sperm viability, AR and mitochondrial function. We conclude that sperm at advanced stages of spermatogenesis, through its passage and storage in the epididymis compartments as well as in the ejaculate, is sensitive to herbicide. The results suggest that ATZ- or DACT-induced disruptions of sperm membranes might impair sperm fertilization competence.

Individual and joint toxicity of the herbicide S-metolachlor and a metabolite, deethylatrazine on aquatic crustaceans: Difference between ecological groups.

We studied the individual and joint acute toxicity of S-metolachlor (SMOC) and deethylatrazine (DEA - a metabolite of atrazine) on different non-target freshwater crustaceans. We used animals from different ecological groups: two amphipods from surface running water (Gammarus pulex and Gammarus cf. orinos), an isopod from surface stagnant water (Asellus aquaticus) and an amphipod living in groundwater (Niphargus rhenorhodanensis). Organisms were exposed to different levels of SMOC and DEA, alone or in binary mixture. Temperature effect on SMOC toxicity was assessed by exposing G. pulex and N. rhenorhodanensis to SMOC at 11 °C and 15 °C. Studying mortality as the biological endpoint, N. rhenorhodanensis was more resistant than surface water species towards SMOC and DEA. Among surface water species, G. pulex was the most sensitive while Gammarus cf. orinos and A. aquaticus showed similar responses to both compounds. Temperature increase did not change SMOC toxicity but modify the shape and steepness of the dose-response curve. We used a Model Deviation Ratio (MDR) approach to evaluate the predictability of Concentration Addition (CA) and Independent Action (IA) models to mixture toxicity. Results indicated either an additive or an antagonistic or a synergistic interaction depending on the concentrations combination and the test species. Our finding conclusively show the suitability of CA and IA in predicting mixture toxicities but results should be interpreted with caution according to ecological group of exposed species in risk assessment procedures.

Kinetics of atrazine, deisopropylatrazine, and deethylatrazine soil biodecomposers.

Twenty-two experimental sets were used to determine the biodecomposition parameters of atrazine (ATZ), deisopropylatrazine (DIATZ), and deethylatrazine (DEATZ) by inverse solution of Michaelis-Menten-Monod kinetic equations. The averaged maximum specific growth rate (µ), Michaelis-Menten half-saturation concentration (K), and biomass yield (Y) ranged between 2.00 × 10-7 and 4.62 × 10-5 1/s, 3.43 × 10-6 and 1.39 × 101 mol/L, and 1.20 × 102 and 2.98 × 105 mg-wet-Bio/mol-Subs, respectively. Parameters grouped by reaction pathway appeared clustered by aerobic and anaerobic catabolic breakdown, and were poorly correlated between each other (R ranging from -0.27 to 0.63, p ≥ 0.05). The tested bacterial strains decomposed ATZ, DIATZ, and DEATZ relatively rapidly in laboratory conditions, with an half-life (t1/2) ranging between 3 and 6 days. Numerical modeling showed that ATZ, DIATZ, and DEATZ half-lives were particularly sensitive to their initial concentration and the initial microbial biomass concentration. This study suggests that these bacterial strains can effectively be used or enhanced for bioremediation of agricultural soils where atrazine has been applied as long as these bacteria already coexist in or can integrate with the local soil microbial population at a given location.

Atrazine and its metabolites degradation in mineral salts medium and soil using an enrichment culture.

An atrazine-degrading enrichment culture was used to study degradation of atrazine metabolites viz. hydroxyatrazine, deethylatrazine, and deisopropylatrazine in mineral salts medium. Results suggested that the enrichment culture was able to degrade only hydroxyatrazine, and it was used as the sole source of carbon and nitrogen. Hydroxyatrazine degradation slowed down when sucrose and/or ammonium hydrogen phosphate were supplemented as the additional sources of carbon and nitrogen, respectively. The enrichment culture could degrade high concentrations of atrazine (up to 110 µg/mL) in mineral salts medium, and neutral pH was optimum for atrazine degradation. Further, except in an acidic soil, enrichment culture was able to degrade atrazine in three soil types having different physico-chemical properties. Raising the pH of acidic soil to neutral or alkaline enabled the enrichment culture to degrade atrazine suggesting that acidic pH inhibited atrazine-degrading ability. The study suggested that the enrichment culture can be successfully utilized to achieve complete degradation of atrazine and its persistent metabolite hydroxyatrazine in the contaminated soil and water.

Degradation of atrazine by UV/chlorine: Efficiency, influencing factors, and products.

In this work, the degradation of atrazine by the combination of UV and chlorine (UV/chlorine) due to the formation of radicals during chlorine photolysis was systematically investigated in terms of efficiency, factors that influence the degradation kinetics, as well as oxidation products. It was found that the degradation efficiency of atrazine was enhanced by UV/chlorine compared to UV or chlorine alone. The degradation efficiency of atrazine was favorable at a lower pH, but was inhibited in the presence of natural organic matters. Meanwhile, the initial chlorine dosage, alkalinity, and chloride barely influenced the degradation efficiency under neutral pH conditions. The degradation of atrazine by UV/chlorine was inhibited in real waters (i.e., surface water and ground water) compared to in deionized water but was still more effective than UV alone. The oxidation products of atrazine resulting from de-alkylation, dechlorination-hydroxylation, alkylic-hydroxylation, alkylic-oxidation, alkylic-hydroxylation-dehydration, deamination-hydroxylation, and dechlorination-hydrogenation in UV/chlorine process were detected, which were slightly different from those formed in UV/H2O2 (commonly used UV-based advanced oxidation process). Particularly, the yields of three primary transformation products (desethyl-atrazine (DEA), desisopropyl-atrazine (DIA), and desethyl-desisopropyl-atrazine (DEIA)) were comparatively quantified in these two processes. The different trend of them formed in UV/chlorine system (DEA:DIA≈4) compared to that formed in UV/H2O2 system (DEA:DIA≈1) could be ascribed to the different reaction reactivities and mechanisms between HO• and Cl• with atrazine.

Atrazine and its main metabolites alter the locomotor activity of larval zebrafish (Danio rerio).

Atrazine (ATZ) and its main chlorometabolites, i.e., diaminochlorotriazine (DACT), deisopropylatrazine (DIP), and deethylatrazine (DE), have been widely detected in aquatic systems near agricultural fields. However, their possible effects on aquatic animals are still not fully understood. In this study, it was observed that several developmental endpoints such as the heart beat, hatchability, and morphological abnormalities were influenced by ATZ and its metabolites in different developmental stages. In addition, after 5 days of exposure to 30, 100, 300 µg L(-1) ATZ and its main chlorometabolites, the swimming behaviors of larval zebrafish were significantly disturbed, and the acetylcholinesterase (AChE) activities were consistently inhibited. Our results also demonstrate that ATZ and its main chlorometabolites are neuroendocrine disruptors that impact the expression of neurotoxicity-related genes such as Ache, Gap43, Gfap, Syn2a, Shha, Mbp, Elavl3, Nestin and Ngn1 in early developmental stages of zebrafish. According to our results, it is possible that not only ATZ but also its metabolites (DACT, DIP and DE) have the same or even more toxic effects on different endpoints of the early developmental stages of zebrafish.