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Atrazine (ATZ), an herbicide widely distributed on a global scale, possess a potential risk for the development of various cancers upon environmental exposure. However, the effect and molecular mechanism of ATZ in cholangiocarcinoma (CCA), is still unclear. This study aimed to investigate the effect of ATZ on the proliferation and migration of CCA cell in vitro. Immortalized human cholangiocytes (MMNK-1) and three CCA cell lines (KKU-055, KKU-100 and KKU-213B) were treated with 0.01 to 100 μM of ATZ and 17β-estradiol (E2). The results showed that, similar to E2, low doses (0.01 to 1 μM) of ATZ promoted the proliferation of all CCA and MMNK-1 cells. ATZ exposure increased non-genomic G protein-coupled estrogen receptor (GPER) expression in the cell membrane and cytoplasm of KKU-213B and KKU-055 cells via G2/M cell cycle accumulation. This, in turn, promoted the proliferation and migration of CCA cells. ATZ exposure induced the upregulation of GPER and increased expression levels of PI3K, p-PI3K, Akt, p-Akt, NF-κB and PCNA. In contrast, following ATZ treatment, the GPER antagonist G15 significantly downregulated the GPER/PI3K/Akt/NF-κB pathway. These results suggest that ATZ promotes CCA cell proliferation and migration through the GPER/PI3K/Akt/NF-κB pathway. This information can enhance public health awareness regarding ATZ contamination to prevent the relative risk of CCA., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work report in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

In planta expression of recombinant antibodies has been proposed as a strategy for herbicide resistance but is not well advanced yet. Here, an atrazine nanobody gene fused with a green fluorescent protein tag was transformed to Arabidopsis thaliana , which was confirmed with PCR, ELISA, and immunoblotting. High levels of nanobody accumulation were observed in the nucleus, cytoderm, and cytosol. The nanobody expressed in the plant had similar affinity, sensitivity, and selectivity as that expressed in Escherichia coli . The T3 homozygous line showed resistance in a dose-dependent manner up to 380 g ai/ha of atrazine, which is approximately one-third of the recommended field application rate. This is the first report of utilizing a nanobody in plants against herbicides. The results suggest that utilizing a high-affinity herbicide nanobody gene rather than increasing the expression of nanobodies in plants may be a technically viable approach to acquire commercial herbicide-resistant crops and could be a useful tool to study plant physiology.

In Brief: Atrazine, like oestrogen, disorganises laminin formation and reduces the number of germ cells and Sertoli cells in the developing testes of the tammar wallaby. This study suggests that interfering with the balance of androgen and oestrogen affects the integrity of laminin structure and testis differentiation., Abstract: The herbicide atrazine was banned in Europe in 2003 due to its endocrine disrupting activity but remains widely used. The integrity of the laminin structure in fetal testis cords requires oestrogen signalling but overexposure to xenoestrogens in the adult can cause testicular dysgenesis. However, whether xenoestrogens affect laminin formation in developing testes has not been investigated. Here we examined the effects of atrazine in the marsupial tammar wallaby during early development and compare it with the effects of the anti-androgen flutamide, oestrogen, and the oestrogen degrader fulvestrant. The tammar, like all marsupials, gives birth to altricial young, allowing direct treatment of the developing young during the male programming window (day 20-40 post partum (pp)). Male pouch young were treated orally with atrazine (5 mg/kg), flutamide (10 mg/kg), 17β-oestradiol (2.5 mg/kg) and fulvestrant (1 mg/kg) daily from day 20 to 40 pp. Distribution of laminin, vimentin, SOX9 and DDX4, cell proliferation and mRNA expression of SRY, SOX9, AMH, and SF1 were examined in testes at day 50 post partum after the treatment. Direct exposure to atrazine, flutamide, 17β-oestradiol, and fulvestrant all disorganised laminin but had no effect on vimentin distribution in testes. Atrazine reduced the number of germ cells and Sertoli cells when examined at day 40-50 pp and day 20 to 40 pp, respectively. Both flutamide and fulvestrant reduced the number of germ cells and Sertoli cells. Atrazine also downregulated SRY expression and impaired SOX9 nuclear translocation. Our results demonstrate that atrazine can compromise normal testicular differentiation during the critical male programming window.

Atrazine (ATZ) is a widely used herbicide that has toxic effects on animals. Melatonin (MLT) is a natural hormone with strong antioxidant properties. However, the effect of MLT on the glucose metabolism disorder caused by ATZ is still unclear. Mice were divided into four groups randomly and given 21 days of gavage: blank control group (Con), 5 mg/kg MLT group (MLT), 170 mg/kg ATZ group (ATZ), and 170 mg/kg ATZ and 5 mg/kg MLT group (ATZ + MLT). The results show that ATZ alters mRNA levels of metabolic enzymes related to glycogen synthesis and glycolysis and increased metabolites (glycogen, lactate, and pyruvate). ATZ causes abnormalities in glucose metabolism in mouse liver, interfering with glycemia regulation ability. MLT can regulate the endoplasmic reticulum to respond to disordered glucose metabolism in mice liver. This study suggested that MLT has the power to alleviate the ATZ-induced glycogen overdeposition and glycolytic deficit.

A common herbicide, atrazine, is associated with poor health. Atrazine acts as an endocrine disruptor at supra-environmental levels. Little research, however, has been conducted regarding chronic exposure to environmental atrazine concentrations across generations. This study utilized comprehensive endpoint measures to investigate the effects of chronic exposure to a conservative atrazine concentration (0.02 ng/mL), measured in Australian waterways, on male mice fertility across two generations. Mice were exposed through the maternal line, from the pre-conception period and through the F1 and F2 generations until three or six months of age. Atrazine did not impact sperm function, testicular morphology nor germ cell parameters but did alter the expression of steroidogenic genes in the F1, down-regulating the expression of Cyp17a1 (Cytochrome P450 family 17, subfamily A member 1; p = 0.0008) and Ddx4 (DEAD-box helicase 4; p = 0.007), and up-regulating the expression of Star (Steroidogenic acute regulatory protein; p = 0.017). In the F2, atrazine induced up-regulation in the expression of Star ( p = 0.016). The current study demonstrates that chronic exposure to an environmentally relevant atrazine concentration perturbs testicular steroid-associated gene expression that varies across generations. Future studies through the paternal and combined parental lineages should be undertaken to further elucidate the multigenerational effects of atrazine on male fertility.

Aquatic organisms are exposed to chemical pesticides including glyphosate (Sharp 480 SL) and atrazine (Atraforce), two phytocidal molecules used for agriculture purposes in Benin. In this study, we assessed the acute toxicity of these two herbicides with emphasis on their histopathological effects on the liver of catfish Clarias gariepinus. One hundred and eighty juveniles of C. gariepinus (mean length 7.26 ± 0.59 cm and mean weight 5.21 ± 3.22 g) were exposed over 96 h to increasing concentrations of each phytocide. The values of 96 h-LC 50 were 6.175 × 10 3 and 3.165 ppm, respectively for Sharp 480 SL and Atraforce. This indicates that Sharp 480 SL was nontoxic, while Atraforce displayed a moderate toxicity to C. gariepinus juveniles. During the tests, the behavioral responses (hyperexcitation, lethargy, loss of balance, discoloration of skin, etc.) that usually precede death were observed in exposed fishes, confirming the neurotoxicity of these phytocides. Histological alterations observed in liver of contaminated fishes were regressive changes, such as necrosis, hepatocyte vacuolation, nuclear degeneration, hepatocytes degeneration, sinusoids dilatation, etc. These results indicate that exposure to these herbicides had destructive effects on the liver of C. gariepinus .

One of many noteworthy consequences of increasing societal reliance on pesticides is their predominance in aquatic environments. These pernicious chemicals interact with high temperatures from global climate change, heat waves, and natural variations to create unstable environments that negatively impact organisms' health. To understand these conditions, we examined the dose-dependent effects of environmentally relevant pesticide mixtures (metolachlor, linuron, isoproturon, tebuconazole, aclonifen, atrazine, pendimethalin, and azinphos-methyl) combined with elevated temperatures (22 control vs. 32°C for 4-week exposure) on renin, dinitrophenyl protein (DNP, an indicator of reactive oxygen species, ROS), 3-nitrotyrosine protein (NTP, an indicator of reactive nitrogen species, RNS), superoxidase dismutase (SOD, an antioxidant), and catalase (CAT, an antioxidant) expressions in the kidneys of goldfish (Carassius auratus). Histopathological analysis showed widespread damage to kidney tissues in high temperature and pesticide co-exposure groups, including rupture of the epithelial layer, hemorrhaging, and degeneration of tubular epithelium. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemical analyses demonstrated significant declines in renin receptor-like mRNA and protein expressions in kidney tissues under combined exposure to high temperature and pesticides compared with controls; conversely, expression of DNP, NTP, SOD, and CAT increased in kidney tissues under the same conditions. Apoptotic cells were also increased in co-exposure groups as assessed by in situ terminal deoxynucleotidyl transferase dUTP nick labeling (TUNEL) assay. The enhanced apoptosis in kidneys of heat and pesticides co-exposed fish was associated with increased caspase-3 (a protease enzyme) mRNA levels. Our results demonstrated that high temperature and pesticides induced oxidative/nitrative stress (i.e., ROS/RNS), damaged tissues, increased cellular apoptosis, and suppressed renin expression in kidneys of goldfish., (© 2022 John Wiley & Sons Ltd.)

Background: The efficacy of a herbicide as soil treatment agent may be largely affected by soil characteristics. Understanding the relationship between herbicide efficacy and soil characteristics can provide decision basis for herbicide application according to local conditions. This study was aimed towards exploring the effect of soil characteristics on herbicidal activity of atrazine as a model herbicide to barnyard grass and thus to find an indicator for the herbicidal activity assessment of a herbicide against weeds., Results: The herbicidal activity of atrazine to barnyard grass varied greatly among the tested soils with the medium inhibition concentration (IC 50 ), based on the amended concentration, ranging from 1.07 to 10.91 mg kg -1 . Uptake of atrazine by barnyard grass was negatively correlated with its adsorption onto soils, whereas it was positively related to the concentration of the herbicide in in situ pore water (C IPW ). Comparable IC 50 values ranging from 1.14 to 1.38 were obtained from C IPW in the tested soils with much smaller variation coefficient compared to those based on the traditional concentration (C soil ) of this herbicide in soils determined by extraction with organic solvents., Conclusion: The concentration of atrazine in in situ pore water could be reliable to evaluate its bioavailability and herbicidal activity to barnyard grass. C IPW of a herbicide in soil could be an indicator for guiding the practical application rate. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Poly(epsilon-caprolactone) nanoparticles are an efficient carrier system for atrazine. However, there is a gap regarding the effects of nanoencapsulation on herbicide-plant interactions. Here, we evaluate the fate and photosystem II inhibition of nano and commercial atrazine in hydroponically grown mustard ( Brassica juncea ) plants whose roots were exposed to the formulations. In addition, to quantify the endogenous levels of atrazine in plant organs, we measured the inhibition of photosystem II activity by both formulations. Moreover, the fluorescently labeled nanoatrazine was tracked in plant tissues using confocal microscopy. The nanoencapsulation induced greater inhibition of photosystem II activity as well as higher accumulation of atrazine in roots and leaves. The nanoparticles were quickly absorbed by the roots, being detected in the vascular tissues and the leaves. Overall, these results provide insights into the mechanisms involved in the enhanced preemergent herbicidal activity of nanoatrazine against target plants.

A three-dimensional (3D) cell-based electrochemical sensor was developed to evaluate the immunomodulatory effects of atrazine and its metabolites by monitoring nitric oxide (NO) release. Nafion/Fe(III) meso-tetra (4-carboxyphenyl) porphyrin/reduced graphene oxide (Nafion/FeTCP@RGO) was functionalized on the screen-printed carbon electrode (SPCE) as a working electrode to enhance the NO selectivity and sensitivity. RAW264.7 cells encapsulated in a gelatin methacrylate (GelMA) hydrogel, forming a 3D cell culture system, were immobilized on the Nafion/FeTCP@RGO/SPCE to serve as biorecognition elements. The proposed sensor presented the induced NO expression as a distinct single differential pulse voltammetric (DPV) anodic peak in the selected physiological lipopolysaccharide (LPS) concentration range (0.01-2000 ng/mL). Furthermore, the peak current intensity of the NO was linearly correlated with the LPS concentration logarithm, which was further validated using a Griess reagent kit. The proposed sensor was utilized in optimized LPS-induction conditions to compare the immunosuppression effect of atrazine with its main metabolites (i.e. desethylatrazine (DEA), deisopropylatrazine (DIA), and diaminochlorotriazine (DACT)). The results demonstrated that the order of 30% inhibitory concentrations was atrazine (25.71 ± 1.08 μg/mL) < DEA (48.63 ± 2.17 μg/mL) ≤ DACT (49.11 ± 1.98 μg/mL) ≤ DIA (52.36 ± 2.34 μg/mL). Overall, this work provided a potential in vitro approach to evaluate the immunotoxicity of pesticides and their metabolites., (Copyright © 2022 Elsevier B.V. All rights reserved.)

It is proposed that the herbicide terbuthylazine is more effective than atrazine in controlling weeds in maize. This study aimed to evaluate the efficacy of terbuthylazine and atrazine in a mixture with glyphosate in glyphosate-tolerant maize for post-emergence application. The experiment was conducted over three trials using randomized blocks with 4 repetitions and 10 treatments, composed by terbuthylazine rates + glyphosate, atrazine rates + glyphosate, [atrazine + mesotrione] + glyphosate, atrazine + tembotrione, isolated glyphosate, and nontreated control. Trial 1 were infested with Bidens subalternans DC. and Commelina benghalensis L; trial 2 with Urochloa plantaginea (Link) R. D. Webster, Ipomoea spp., volunteer soybean, B. subalternans, and grasses; and trial 3 infestation with C. benghalensis , U. plantaginea , Ipomoea spp., volunteer soybean, B. subalternans , Amaranthus hybridus L., and grasses. Weed control, crop injury, and yield were evaluated. Terbuthylazine + glyphosate showed an efficacy equivalent to that of atrazine or [atrazine + mesotrione] + glyphosate in the control of broadleaves and C. benghalensis . In contrast, the efficacy of terbuthylazine was similar or greater than that observed for atrazine in controlling grasses, depending on the location. Terbuthylazine is an important partner of glyphosate in controlling weeds in maize and is an alternative to atrazine.

Background/aims: Pulmonary fibrosis can be caused by genetic abnormalities, autoimmune disorders or exposure to environmental pollutants. All these causes have in common the excessive production of oxidative stress species that initiate a cascade of molecular mechanism underlying fibrosis in a variety of organs, including lungs. The chemical name of Atrazine (ATR) is 6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4-diamine, and it is the most commonly used broad-spectrum herbicide in agricultural crops. Additionally, Bleomycin is a chemotherapeutic agent often used for different lymphoma with a seriously pulmonary complication. The most accredited hypothesis that may explain the mechanism of toxicity induced by ATR or bleomycin is exactly the production of reactive oxygen species (ROS) that leads to an unbalance in the physiological anti-oxidant system. However, until today, nobody has investigated the effect of ATR exposure during pulmonary fibrosis., Methods: Mice were subject to ATR exposure, to bleomycin injection or to both. At the end of experiment, the lungs and blood were collected. Additionally, we analyzed by different test such as open field, pole and rotarod test or other we investigated the effects of ATR or bleomycin exposure on behavior., Results: Following ATR or bleomycin induction, we found a significant increase in lung damage, fibrosis, and oxidative stress. This condition was significantly worsened when the animals injected with bleomycin were also exposed to ATR. Additionally, we observed significant motor and non-motor impairment in animals exposed to ATR., Conclusion: Our study demonstrates that ATR exposure, decrease nuclear factor-erythroid 2-related factor (Nrf2) pathways in both lung and brain., Competing Interests: The authors have no conflicts of interest to declare., (© Copyright by the Author(s). Published by Cell Physiol Biochem Press.)

The rapid development of nanotechnology influences the developments within the agro-sector. An example is provided by the production of nanoenabled pesticides with the intention to optimize the efficiency of the pesticides. At the same time, it is important to collect information on the unintended and unwanted adverse effects of emerging nanopesticides on nontarget plants. Currently, this information is limited. In the present study, we compared the effects of a nanoformulation of atrazine (NPATZ) and the nonencapsulated atrazine formulation (ATZ) on physiological responses, defense mechanisms, and nutrient displacement in lettuce over time with the applied concentrations ranging from 0.3 to 3 mg atrazine per kg soil. Our results revealed that both NPATZ and ATZ induced significant decreases in plant biomass, chlorophyll content, and protein content. Additionally, exposure to NPATZ and ATZ caused oxidative stress to the lettuce plant and significantly elevated the activities of the tested ROS scavenger enzymes in plant tissues. These results indicate that NPATZ and ATZ cause distinct adverse impacts on lettuce plants. When comparing the adverse effects in plants after exposure to NPATZ and ATZ, no obvious differences in plant biomass and chlorophyll content were observed between NPATZ and ATZ treatments at the same exposure concentration regardless of exposure duration. An enhanced efficiency of the active ingredient of the nanopesticide as compared to the conventional formulation was observed after long-term exposure to the high concentration of NPATZ, as it induced higher impacts on plants in terms of the end points of the contents of protein, superoxide anion (O 2 ̇ - ), and MDA, and the activities of stress-related enzymes as compared to the same concentration of ATZ. Furthermore, exposure to both NPATZ and ATZ disrupted the uptake of mineral nutrients in plants, and the differences in the displacement of nutrients between the NPATZ and ATZ treatments depended on the element type, plant organ, exposure concentration, and time. Overall, the application dose of a nanopesticide should balance their increased herbicidal efficiency with the long-term adverse effects in order to maximize the desired impact while minimizing adverse impacts; only then will we be able to understand the potential impact of nanopesticides on the environment.

Background: exposure to environmental contaminants has been linked to an increased risk of neurological diseases and poor outcomes. Chemical name of Atrazine (ATR) is 6-chloro- N -ethyl- N '-(1-methylethyl)-1,3,5-triazine-2,4-diamine, and it is the most commonly used broad-spectrum herbicide in agricultural crops. Several studies have demonstrated that ATR has the potential to be harmful to the brain's neuronal circuits. Until today nobody has explored the effect of ATR inhalation on young and aged mice., Methods: young and aged mice were subject to 25 mg of ATR in a vehicle made with saline and 10% of Dimethyl sulfoxide (DMSO) every day for 28 days. At the end of experiment different behavioral test were made and brain was collected., Results: exposure to ATR induced the same response in terms of behavioral alterations and motor and memory impairment in mice but in aged group was more marked. Additionally, in both young and aged mice ATR inhalations induced oxidative stress with impairment in physiological antioxidant response, lipid peroxidation, nuclear factor kappa-light-chain-enhancer of activated B cells (nf-κb) pathways activation with consequences of pro-inflammatory cytokines release and apoptosis. However, the older group was shown to be more sensitive to ATR inhalation., Conclusions: our results showed that aged mice were more susceptible compared to young mice to air pollutants exposure, put in place a minor physiologically response was seen when exposed to it.

Atrazine is one of the most commonly used pre-emergence and early post-emergence herbicides in the world. We have shown previously that atrazine does not directly stimulate the pituitary or adrenal to trigger hormone release but acts centrally to activate a stress-like activation of the hypothalamic-pituitary-adrenal axis. In doing so, atrazine treatment has been shown to cause adrenal morphology changes characteristic of repeated stress. In this study, adrenals from atrazine treated and stressed animals were directly compared after 4 days of atrazine treatment or restraint stress. Both atrazine and stressed animals displayed reduced adrenocortical zona glomerulosa thickness and aldosterone synthase (CYP11B2) expression, indicative of repeated adrenal stimulation by adrenocorticotropic hormone. To determine if reduced CYP11B2 expression resulted in attenuated aldosterone synthesis, stressed and atrazine treated animals were challenged with angiotensin II (Ang II). As predicted, stressed animals produced less aldosterone compared to control animals when stimulated. However, atrazine treated animals had higher circulating aldosterone concentrations compared to both stressed and control groups. Ang II-induced aldosterone release was also potentiated in atrazine pretreated human adrenocortical carcinoma cells (H295R). Atrazine pretreated did not alter the expression of the rate limiting steroidogenic StAR protein or angiotensin II receptor 1. Atrazine treated animals also presented with higher basal blood pressure than vehicle treated control animals suggesting sustained elevations in circulating aldosterone levels. Our results demonstrate that treatment with the widely used herbicide, atrazine, directly increases stimulated production of aldosterone in adrenocortical cells independent of expression changes to rate limiting steroidogenic enzymes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zimmerman, Mackay, Kemppainen, Jones, Read, Schwartz and Foradori.)

Biomimetic design represents an emerging field for improving knowledge of natural molecules, as well as to project novel artificial tools with specific functions for biosensing. Effective strategies have been exploited to design artificial bioreceptors, taking inspiration from complex supramolecular assemblies. Among them, size-minimization strategy sounds promising to provide bioreceptors with tuned sensitivity, stability, and selectivity, through the ad hoc manipulation of chemical species at the molecular scale. Herein, a novel biomimetic peptide enabling herbicide binding was designed bioinspired to the D1 protein of the Photosystem II of the green alga Chlamydomonas reinhardtii. The D1 protein portion corresponding to the Q B plastoquinone binding niche is capable of interacting with photosynthetic herbicides. A 50-mer peptide in the region of D1 protein from the residue 211 to 280 was designed in silico, and molecular dynamic simulations were performed alone and in complex with atrazine. An equilibrated structure was obtained with a stable pocked for atrazine binding by three H-bonds with SER222, ASN247, and HIS272 residues. Computational data were confirmed by fluorescence spectroscopy and circular dichroism on the peptide obtained by automated synthesis. Atrazine binding at nanomolar concentrations was followed by fluorescence spectroscopy, highlighting peptide suitability for optical sensing of herbicides at safety limits., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Target of rapamycin (TOR) kinase is a sensor and a central integrator of internal and external metabolic cues. However, in algae and in higher plants, the components of TOR kinase signaling are yet to be characterized. Here, we establish an assay system to study TOR kinase activity in Chlamydomonas reinhardtii using the phosphorylation status of its putative downstream target, CrS6K. Using this assay, we probe the modulation of cellular TOR kinase activity under various physiological states such as photoautotrophy, heterotrophy, mixotrophy, and nitrogen (N) starvation. Importantly, we uncover that excess acetate in the medium leads to high cellular reactive oxygen species levels, triggering autophagy and a concomitant drop in TOR kinase activity in a dose-dependent manner, thus leading to a N-starvation-like cellular phenotype, even when nitrogen is present., (© 2020 Federation of European Biochemical Societies.)

Background: Atrazine is one of the most widespread chlorinated herbicides, leaving large bulks in soils and groundwater. The biodegradation of atrazine by bacteria is well described, but many aspects of the fungal metabolism of this compound remain unclear. Thus, we investigated the toxicity and degradation of atrazine by 13 rainforest basidiomycete strains., Results: In liquid medium, Pluteus cubensis SXS320, Gloelophyllum striatum MCA7, and Agaricales MCA17 removed 30, 37, and 38%, respectively, of initial 25 mg L - 1 of the herbicide within 20 days. Deficiency of nitrogen drove atrazine degradation by Pluteus cubensis SXS320; this strain removed 30% of atrazine within 20 days in a culture medium with 2.5 mM of N, raising three metabolites; in a medium with 25 mM of N, only 21% of initial atrazine were removed after 40 days, and two metabolites appeared in culture extracts. This is the first report of such different outcomes linked to nitrogen availability during the biodegradation of atrazine by basidiomycetes. The herbicide also induced synthesis and secretion of extracellular laccases by Datronia caperata MCA5, Pycnoporus sanguineus MCA16, and Polyporus tenuiculus MCA11. Laccase levels produced by of P. tenuiculus MCA11 were 13.3-fold superior in the contaminated medium than in control; the possible role of this enzyme on atrazine biodegradation was evaluated, considering the strong induction and the removal of 13.9% of the herbicide in vivo. Although 88% of initial laccase activity remained after 6 h, no evidence of in vitro degradation was observed, even though ABTS was present as mediator., Conclusions: This study revealed a high potential for atrazine biodegradation among tropical basidiomycete strains. Further investigations, focusing on less explored ligninolytic enzymes and cell-bound mechanisms, could enlighten key aspects of the atrazine fungal metabolism and the role of the nitrogen in the process.

Long-standing evidence supports the importance of maintaining healthy populations of microbiota for the survival, homeostasis, and complete development of marine mollusks. However, the long-term ecological effects of agricultural runoff on these populations remains largely unknown. Atrazine (6-Chloro-n-ethyl-n'-(1-methylethyl)-triazine-2,4-diamine), a prevalent herbicide in the United States, is often used along tributaries of the Chesapeake Bay where oyster breeding programs are concentrated. To investigate any potential effects atrazine maybe having on mollusk-prokaryote interactions, we used 16S rRNA gene amplicons to evaluate how microbial compositions shift in response to exposure of environmentally relevant concentrations of atrazine previously found within the Chesapeake Bay. The dominant bacterial genera found within all groups included those belonging to Pseudoalteromonas, Burkholderia, Bacteroides, Lactobacillis, Acetobacter, Allobaculum, Ruminococcus, and Nocardia. Our results support previously published findings of a possible core microbial community in Crassostrea virginica. We also report a novel finding: oysters exposed to atrazine concentrations as low as 3 µg/L saw a significant loss of a key mutualistic microbial species and a subsequent colonization of a pathogenic bacteria Nocardia. We conclude that exposure to atrazine in the Chesapeake Bay may be contributing to a significant shift in the microbiomes of juvenile oysters that reduces fitness and impedes natural and artificial repopulation of the oyster species within the Bay.

Despite the important ecological and agricultural production value of fallow field vegetation in agricultural landscapes, it is often affected by herbicide drift and runoff from neighboring sprayed fields. However, little is known about the impact of herbicides on the non-target plant community of fallow fields. In this study, the plant community of fallow fields was investigated following annual sublethal exposure to atrazine or tribenuron-methyl by a 3-year (2014-2016) randomized block field study. The two herbicides both changed the species composition, reduced the number of plant species and the relative frequencies of some plants, and significantly reduced the Margalef species richness index and Shannon's diversity index of the plant community in the fallow field. The effects of the two herbicides on species number and community composition were not consistent. The effects of herbicide doses less than the recommended field application concentration (RFAC) on the plant community composition and community diversity of the fallow field were not lower than the effects of the RFAC of the herbicides. Indeed, doses less than the RFAC had an even greater impact on the community diversity than the RFAC of the herbicides. As the number of years of herbicide application increased, the effects of the herbicides on the plant community diversity did not increase compared to the effects of the blank control, and the herbicides did not change the functional composition of the plant communities in the fallow field. Our results suggest that the ecological risks of herbicides, even at low concentrations, on non-target wild plant communities in agricultural landscapes should not be neglected in the development of practical plant diversity conservation strategies.

Single cell lipid profiling is a powerful tool to connect membrane composition and its changes within individual cells to specific biochemical functions or stimuli, but current approaches are inadequate due to the complex nature of the cells and technical limitation in analysis. Herein we report a new method with plasmonic substrates capable of cell localization and enhanced lipid ionization through thin-gold-film MALDI-MS. We performed lipidomic profiling of algae single cells with a 120-well microarray and identified more than 50 lipids in C. reinhardtii without an extraction process. The substrate was used for probing toxicological effect of herbicide atrazine on the algae's lipidome, demonstrating molecular changes in glycerol lipid profiles. Fast location of cells with metal-enhanced fluorescence (MEF) and subsequent precise and direct ionization of the LDI process contribute to the enhanced performance, allowing for assessment of lipid changes concurrent with atrazine affected populations. This method that combines microarrays, MEF, and MALDI-MS presents an effective platform for lipidomic study of single cells and for environmental toxicity study with microorganisms.

Recent empirical evidence suggests that herbicides have damaging effects on non-target organisms in both natural and semi-natural ecosystems. The African mound building termite, Macrotermes bellicosus, is an important beneficial insect that functions as an ecosystem engineer due to its role in the breakdown of dead and decaying materials. Here, we examined the effects of 2,4-D amine salt (2,4-D) and atrazine based herbicides viz. Vestamine® and Ultrazine® on the survival and locomotion response of M. bellicosus. Worker termites were treated with a range of concentrations of Vestamine® (the recommended concentration: 6.25 ml per 500 ml of water, 0.25- and 0.5-fold below the recommended concentration and distilled water as control) and Ultrazine® (the recommended concentration: 3.75 ml per 500 ml of water, 0.25-, 0.5-, 2.0- and 4-fold of the recommended concentration and distilled water as control) for 24 hours for the mortality test, and allowed to run for 15 seconds for the locomotion trial. All concentrations of both Vestamine® and Ultrazine® were highly toxic to worker termites and mortality increased as the concentration and time after treatment increased. For both herbicides, concentrations far less than the recommended rates caused 100% mortality. The speed of termites was significantly influenced by both Vestamine® and Ultrazine® as termites exposed to all tested concentrations of the herbicides exhibited reduced running speed than the control. These findings suggest that beneficial insects, especially M. bellicosus may experience high mortality (up to 100%) and reduced mobility if they are sprayed upon or come in contact with plant materials that have been freshly sprayed with (less or more than) the recommended concentrations of Vestamine® and Ultrazine®. The findings of our study calls for the reassessment of the usage of 2,4-D and atrazine based herbicides in weed control in termite and other beneficial insect populated habitats., Competing Interests: The authors have declared that no competing interests exist.

Atrazine is a triazine herbicide that is widely used to control broadleaf weeds. Its widespread use over the last 50 years has led to the potential contamination of soils, groundwater, rivers, and lakes. Its main route of complete degradation is via biological means, which is carried out by soil microbiota using a 6-step pathway. The aim of the present study was to investigate whether application of atrazine to soil changes the soil bacterial community. We used 16S rRNA gene sequencing and qPCR to elucidate the microbial community structure and assess the abundance of the atrazine degradation genes atzA, atzD, and trzN in a Brazilian soil. The results obtained showed that the relative abundance of atzA and trzN, encoding triazine-initiating metabolism in Gram-negative and -positive bacteria, respectively, increased in soil during the first weeks following the application of atrazine. In contrast, the abundance of atzD, encoding cyanuric acid amidohydrolase-the fourth step in the pathway-was not related to the atrazine treatment. Moreover, the overall soil bacterial community showed no significant changes after the application of atrazine. Despite this, we observed increases in the relative abundance of bacterial families in the 4 th and 8 th weeks following the atrazine treatment, which may have been related to higher copy numbers of atzA and trzN, in part due to the release of nitrogen from the herbicide. The present results revealed that while the application of atrazine may temporarily increase the quantities of the atzA and trzN genes in a Brazilian Red Latosol soil, it does not lead to significant and long-term changes in the bacterial community structure.

Treatment of Arabidopsis thaliana seedlings with the PSII-inhibiting herbicide atrazine results in xenobiotic and oxidative stress, developmental arrest, induction of senescence and cell death processes. In contrast, exogenous sucrose supply confers a high level of atrazine stress tolerance, in relation with genome-wide modifications of transcript levels and regulation of genes involved in detoxification, defense and repair. However, the regulation mechanisms related to exogenous sucrose, involved in this sucrose-induced tolerance, are largely unknown. Characterization of these mechanisms was carried out through a combination of transcriptomic, metabolic, functional and mutant analysis under different conditions of atrazine exposure. Exogenous sucrose was found to differentially regulate genes involved in polyamine synthesis. ARGININE DECARBOXYLASE ADC1 and ADC2 paralogues, which encode the rate-limiting enzyme (EC 4.1.1.19) of the first step of polyamine biosynthesis, were strongly upregulated by sucrose treatment in the presence of atrazine. Such regulation occurred concomitantly with significant changes of major polyamines (putrescine, spermidine, spermine). Physiological characterization of a mutant affected in ADC activity and exogenous treatments with sucrose, putrescine, spermidine and spermine further showed that modification of polyamine synthesis and of polyamine levels could play adaptive roles in response to atrazine stress, and that putrescine and spermine had antagonistic effects, especially in the presence of sucrose. This interplay between sucrose, putrescine and spermine is discussed in relation with survival and anti-death mechanisms in the context of chemical stress exposure., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

The study reports the effects of an herbicide (atrazine) and a plasticizer (Bisphenol A, BPA) on the transcriptional modulation of a mismatch repair gene (mlh1) and its adverse consequences on female fertility using Drosophila as a model. Through a chemical screen, we show that exposure to atrazine or BPA significantly downregulates mlh1 and the exposed flies had reduced fertility with smaller ovaries having reduced number of mature oocytes and abnormal distribution of ovarian follicles with increased apoptosis in them. These females had increased double-strand breaks as well as reduced synaptonemal complex formation in their ovaries suggesting altered meiotic crossing over. The eggs of these females were defective in their maternal transcripts as well as proteins and consequently, after fertilization, these eggs exhibited abnormal embryonic development. Interestingly, these phenotypes parallel that of mlh1 mutants. Further, exposure of females having reduced Mlh1 levels (mlh1 e00130 /CyO) to atrazine or BPA caused severe defective phenotypes at a higher proportion than normal flies. Our findings reveal the critical role of mlh1 in atrazine and BPA mediated female reproductive toxicity, and opens up a possibility of toxicants affecting female fertility by modulating the MMR genes., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Atrazine is one of the most used herbicides and has been associated with persistent surface and groundwater contamination, and novel formulations derived from nanotechnology can be a potential solution. We used poly(ε-caprolactone) nanoencapsulation of atrazine (NC+ATZ) to develop a highly effective herbicidal formulation. Detailed structural study of interaction between the formulation and Brassica juncea plants was carried out with evaluation of the foliar uptake of nanoatrazine and structural alterations induced in the leaves. Following postemergent treatment, NC+ATZ adhered to the leaf and penetrated mesophyll tissue mainly through the hydathode regions. NC+ATZ was transported directly through the vascular tissue of the leaves and into the cells where it degraded the chloroplasts resulting in herbicidal activity. Nanocarrier systems, such as the one used in this study, have great potential for agricultural applications in terms of maintenance of herbicidal activity at low concentrations and a substantial increase in the herbicidal efficacy.

Atrazine (ATZ) is one of the most widely used herbicides worldwide and is a common contaminant in human drinking water. It disrupts metabolic pathways in plants, and has metabolic and reproductive effects in vertebrates, including humans. Few studies have investigated the effects of exposure to low doses of ATZ, especially during sexual development in males. In this study, we exposed C57BL/6J male mice from weaning for 8 weeks to drinking water containing 0.5mgkg-1 bodyweight (BW) day-1 ATZ, the 'no observed effect' level used by the Australian government, or a 10-fold higher dose (5mgkg-1 BW day-1). Mice treated with the low dose of ATZ showed increased total and cumulative weight gain. At 12 weeks of age, there was a significant increase in the percentage of dead spermatozoa in both ATZ-exposed groups, as well as decreased epididymal sperm motility in the low-dose ATZ group. Significant changes in testis and liver gene expression were also observed following ATZ exposure. These data demonstrate that a low dose of ATZ can perturb metabolic and reproductive characteristics in male mice. A chronic reduction in sperm quality and increased weight gain could have negative consequences on the reproductive capacity of males, and further studies should consider the effects of long-term ATZ exposure on male reproductive health.

Main Conclusion: Detrimental pleiotropic effects of resistance mutation(s) were observed for multiple-resistant phenotypes (resistant to both atrazine and dicamba). The multiple-resistant phenotypes had lower growth rates and less capacity for vegetative growth compared to the phenotypes only resistant to atrazine. The fitness costs that are conferred by herbicide resistance alleles can affect the rate of herbicide resistance evolution within populations. We evaluated the direct fitness costs involved with multiple resistance to dicamba and atrazine (R1 and R2) in Chenopodium album by comparing the performance of multiple-resistant phenotypes to those phenotypes that were only resistant to atrazine (S1 and S2). The R1 and R2 phenotypes were consistently shorter and produced less dry matter than the S1 and S2 phenotypes. The R1 and R2 phenotypes were shown to have lower relative growth rates (RGR) and net assimilation rates (NAR) than the S1 and S2 phenotypes at an early stage of growth. However, there was no significant difference in RGR between the R1 and R2 and, S1 and S2 phenotypes at a later stage of growth, though the R1 and R2 phenotypes still had a lower NAR at this later stage. Further investigations using a neighbouring crop competition approach showed that the R1 and R2 phenotypes were weaker competitors, and exhibited significantly less capacity for vegetative growth compared to the S1 and S2 phenotypes during competition. Overall, the results of this study revealed multiple- resistance to atrazine and dicamba endowed a significant fitness penalty to C. album, and it is possible that the frequency of multiple-resistant individuals would gradually decline once selection pressure from herbicides was discontinued.

Modern transformation and genome editing techniques have shown great success across a broad variety of organisms. However, no study of successfully applied genome editing has been reported in a dinoflagellate despite the first genetic transformation of Symbiodinium being published about 20 years ago. Using an array of different available transformation techniques, we attempted to transform Symbiodinium microadriaticum (CCMP2467), a dinoflagellate symbiont of reef-building corals, with the view to performing subsequent CRISPR-Cas9 mediated genome editing. Plasmid vectors designed for nuclear transformation containing the chloramphenicol resistance gene under the control of the CaMV p35S promoter as well as several putative endogenous promoters were used to test a variety of transformation techniques including biolistics, electroporation and agitation with silicon carbide whiskers. Chloroplast-targeted transformation was attempted using an engineered Symbiodinium chloroplast minicircle encoding a modified PsbA protein expected to confer atrazine resistance. We report that we have been unable to confer chloramphenicol or atrazine resistance on Symbiodinium microadriaticum strain CCMP2467., Competing Interests: The authors have declared that no competing interests exist.

Atrazine is one of the most frequently used herbicides in Argentina for controlling broadleaf weeds and annual grasses. Currently, there is limited information on the impact of triazine herbicides on mycotoxin production and growth parameters of toxigenic fungi in maize. The objective of this study was to evaluate the effect of different concentrations of atrazine on the lag phase prior to growth, the growth rate, and on production of aflatoxin B 1 (AFB 1 ) of Aspergillus flavus and Aspergillus parasiticus strains, on maize meal extract agar (MMEA) under different water activities (a W ) and temperatures. A commercial formulation of atrazine was added to MMEA medium at 0, 5, 10, 50, or 100 mmol/l, adjusted to 0.98, 0.95, and 0.93 a W , and incubated at 28 °C and 37 °C for 21 days. AFB 1 was determined by HPLC after 7, 14, and 21 days of incubation. In the control treatments, a significant increase in the time prior to growth was observed and as the a W decreased, at both temperatures, the growth rate of the strains also decreased. A significant increase in growth rate was observed as the concentration of atrazine in the medium increased, for all a W levels tested. The optimal conditions for the accumulation of AFB 1 in the control treatments were 0.98 a W and 28 °C, after 7 days of incubation. As the concentration of herbicide increased, AFB 1 production also increased (P < 0.05). These results add to the knowledge about consequences with regard to aflatoxin production of the use of excessive atrazine doses in extensive maize culture.

Background: Wild radish (Raphanus raphanistrum) is a globally important weed of crops. Two atrazine-resistant wild radish populations (R1 and R2), collected from the Western Australia grain belt, were investigated for resistance to photosystem II (PSII) herbicides., Results: Sequencing of the full-length psbA gene revealed the well-known Ser264-Gly substitution in population R1, whereas population R2 displayed a novel Phe274-Val substitution. Herbicide dose-response studies confirmed that the population with the Ser264-Gly mutation exhibited high-level resistance to atrazine, but super-sensitivity to bromoxynil. Plants possessing the novel Phe274-Val mutation exhibited a modest level of resistance to atrazine, metribuzin and diuron, and were bromoxynil susceptible. Structural modelling of the mutant D1 proteins predicts that the Ser264-Gly mutation endows atrazine resistance by abolishing H-bonds, but confers bromoxynil super-sensitivity by enhancing hydrogen bonding. The Phe274-Val substitution provides resistance to atrazine and diuron by indirectly affecting H-bond formation between the Ser264 residue and the herbicides., Conclusion: The results demonstrate that the Phe274-Val mutation is likely responsible for resistance to PSII-inhibiting triazine and urea herbicides. To our knowledge, this is the first evidence of the psbA Phe274-Val mutation in wild radish conferring resistance to PSII herbicides. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Atrazine, a commonly used herbicide, suppresses the luteinizing hormone (LH) surge in female rats, although the underlying mechanism remains unclear. Kisspeptin, encoded by the Kiss1 gene, is a hypothalamic peptide that controls gonadotropin-releasing hormone (GnRH) release from the GnRH neurons. Kisspeptin neurons in the anteroventral periventricular nucleus (AVPV) are involved in regulating pre-ovulatory GnRH and LH surge. To clarify the effect of atrazine on the LH surge in female rats, we investigated its effects on hypothalamic GnRH and kisspeptin. Ovariectomized female rats in a high-dose estradiol supplementation model were orally administered vehicle or 100 mg/kg of atrazine once daily for 5 days. This attenuated the LH surge but did not affect baseline LH levels, with no difference in hypothalamic GnRH levels between the vehicle-treated and atrazine-treated animals. After the fifth treatment, subcutaneous administration of kisspeptin (at 0, 0.1, 1, and 10 nmol/kg) induced a dose-dependent LH release almost equivalent in the vehicle- and atrazine-treated animals, suggesting that GnRH neurons maintain normal responsiveness to kisspeptin. However, Kiss1 mRNA expression levels in the AVPV were significantly reduced in the atrazine-treated animals. Given the normal response of GnRH neurons to exogenously administered kisspeptin, the suppressive effect of atrazine may be explained by suppression of Kiss1 expression in the AVPV leading to the attenuation of kisspeptin release from kisspeptin neurons in the AVPV. Further studies are warranted to elucidate more precisely the mechanism of atrazine's involvement in the suppression of Kiss1 mRNA expression in the AVPV.

Amphibians are considered to be excellent bioindicators to their morphophysiological characteristics and life cycle. In this context, the present study investigated the morphological integrity of anuran larvae collected in preserved environments in the Emas National Park, in the municipality of Mineiros (Goiás state, Brazil), and in environments representative of the agricultural matrix of the Rio Verde region, also in Goiás, where there is a long history of the use of agricultural pesticides. Samples of water from temporary ponds, permanent dams, and veredas were analyzed for the presence of pesticides and, especially atrazine (5350 μg/L), found at significantly higher concentrations in the agricultural matrix. We observed a high percentage (approximately 10%) of morphological malformations including alterations of the fins in Boana albopunctatus and Scinax fuscovarius; alteration in oral structures in B. albopunctatus, Dematonotus muelleri, Physalaemus centralis, Physalaemus cuvieri, and Leptodactylus fuscus mainly in the tadpoles collected in the agricultural environment in comparison with those from the protected area (3.5%; P < 0.0001, χ 2 31.75). However, changes in the eyes, mouth, intestines, and nostrils, as well as amelia were observed only in the agricultural environment. The vast majority of the observed malformations were associated with the tail and oral disc, which suggests that these anatomical parameters may be used as sensitive morphological biomarkers. Given these findings, we reinforce that areas of agricultural land may have a deleterious effect on the morphological integrity of the tadpoles and consequently, on their development, and that these features may be used as indicators of environmental quality and health.

This study evaluated the in vitro effect of three concentrations of atrazine, chlorpyrifos and endosulfan on the growth parameters of four non-toxigenic Aspergillus section Flavi strains. The ability of the strains to remove these pesticides in a synthetic medium was also determined. Growth parameters were measured on soil extract solid medium supplied with 5, 10 and 20mg/l of each pesticide, and conditioned to -0.70, -2.78, -7.06 and -10.0 water potential (MPa). Removal assays were performed in Czapek Doc medium (CZD) supplied with 20mg/l of each pesticide under optimal environmental conditions (-2.78 of MPa and 25°C). The residual levels of each pesticide were detected by the reversed-phase HPLC/fluorescence detection system. The lag phases of the strains significantly decreased in the presence of the pesticides with respect to the control media. This result indicates a fast adaptation to the conditions assayed. Similarly, the mycelial growth rates in the different treatments increased depending on pesticide concentrations. Aspergillus oryzae AM 1 and AM 2 strains showed high percentages of atrazine degradation (above 90%), followed by endosulfan (56 and 76%) and chlorpyrifos (50 and 73%) after 30 days of incubation. A significant (p<0.001) correlation (r=0.974) between removal percentages and growth rate was found. This study shows that non-toxigenic Aspergillus section Flavi strains from agricultural soils are able to effectively grow in the presence of high concentrations of atrazine, chlorpyrifos and endosulfan under a wide range of MPa conditions. Moreover, these strains have the ability to remove high levels of these pesticides in vitro in a short time., (Copyright © 2018 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

The extensive use of pesticides represents a risk to human health and to the environment. This study aimed to investigate if the exposure to atrazine and diuron, two herbicides widely used in Brazil, could induce changes in the susceptibility profile to aztreonam, colistin and polymyxin B antimicrobials in isolates of P. aeruginosa obtained from soil samples by using the determination of minimum inhibitory concentration (MIC) test. Three isolates had an increase of MIC to aztreonam after exposure to both herbicides and one isolate did not show any MIC change. The MexAB-OprM efflux pump has already been upregulated in these isolates and the herbicides atrazine and diuron did not increase MexAB-OprM overexpression. Therefore, the decrease in aztreonam susceptibility was not directly related to this pump, suggesting that probably other mechanisms should be involved.

The effects of wheat root exudates and the aging interactions between biochar and soil on atrazine desorption from biochar-amended soil were carefully examined. Compared with CaCl 2 solution, wheat root exudates significantly increase the desorption of atrazine from biochar, mainly by promoting the desorption of atrazine adsorbed on biochar with specific forces. Wheat root exudates were effectively separated into three components with different electrical properties, namely, anionic, neutral, and cationic components. Mainly due to the carboxyl-containing compounds, the anionic component was the main active component in the wheat root exudates that enhances the desorption of atrazine from the biochar. Additionally, wheat root exudates can increase the desorption of atrazine from biochar-amended soil. The promotion of atrazine desorption by root exudates was more obvious in soils with low organic matter contents, where atrazine was mainly adsorbed by biochar. The aging interaction between the biochar and soil increased the total desorption rate and rapid desorbing fraction of the atrazine in the soil, most likely due to the reduction of the biochar sorption capacity in the aged biochar-amended soil., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Herbicides can impact non-target metabolic pathways in natural enemies and lead to the reduction of these populations in the field. Behavioral characteristics, morphology and histology of reproductive structures and reproduction of females of Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) were evaluated under the effect of the herbicides atrazine, nicosulfuron and the mixture of both. The number of mature oocytes per ovary was lower in females exposed to the herbicides atrazine, nicosulfuron and the mixture of both. Herbicides did not affect the longevity and mortality of P. nigrispinus, therefore, they are selective for this predator. On the other hand, herbicides can cause sublethal effects by affecting the reproduction of predators.

Human activities have caused increasing inputs of pharmaceuticals to the environment and diclofenac (DF) is one of the most commonly detected in freshwater systems. The aim of this study was to determine the impact of DF on a freshwater green alga as a non-target organism. For DF toxicity evaluation, its effects on a model organism Chlamydomonas reinhardtii were compared with effects caused by the herbicide atrazine (AT). EC 50 values were about 135 mg/L for DF and 78 mg/L for AT, respectively. Both toxicants enhanced H 2 O 2 production by the cells (144% and 178% of control for AT and DF, respectively) and stimulated catalase activity (≈200% of control). Activity of ascorbate peroxidase was elevated in AT-cells but not in DF-treated cells. DF did not influence dark respiration of the cells, whereas AT inhibited this process by about 50% compared to the control. Both toxicants caused photosynthesis inhibition. Analysis of parameters of chlorophyll a fluorescence in vivo showed diminishment of a performance index (PI) in both DF- and AT-treated cells (≈50% of control), but the reasons for the changes detected were different. AT diminished the efficiency of electron transport between PS II and PS I without significant inhibition of PS II or PS I reaction centers (RCs). In contrast to AT, DF seemed to influence directly PS II RCs. The fraction of active PS II RCs was lowered in DF-treated cells, but energy flux per active RC increased. Our study indicates that DF phytotoxicity results mainly from photosynthesis inhibition due to "silencing" of a fraction of PS II RCs., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

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.

The extent of residual contaminations of pesticides through drift, run-off and leaching is a potential threat to non-target plant communities. Arabidopsis thaliana responds to low doses of the herbicide atrazine, and of its degradation products, desethylatrazine and hydroxyatrazine, not only in the long term, but also under conditions of short-term exposure. In order to investigate underlying molecular mechanisms of low-dose responses and to decipher commonalities and specificities between different chemical treatments, parallel transcriptomic studies of the early effects of the atrazine-desethylatrazine-hydroxyatrazine chemical series were undertaken using whole-genome microarrays. All of the triazines under study produced coordinated and specific changes in gene expression. Hydroxyatrazine-responsive genes were mainly linked to root development, whereas atrazine and desethylatrazine mostly affected molecular signaling networks implicated in stress and hormone responses. Analysis of signaling-related genes, promoter sites and shared-function interaction networks highlighted the involvement of energy-, stress-, abscisic acid- and cytokinin-regulated processes, and emphasized the importance of cold-, heat- and drought-related signaling in the perception of low doses of triazines. These links between low-dose xenobiotic impacts and stress-hormone crosstalk pathways give novel insights into plant-pesticide interactions and plant-pollution interactions that are essential for toxicity evaluation in the context of environmental risk assessment., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Atrazine (ATZ), the second most commonly used herbicide in the United States, is an endocrine disrupting chemical linked to cancer and a common drinking water contaminant. This study further investigates ATZ-related developmental toxicity by testing the following hypotheses in zebrafish: the effects of embryonic ATZ exposure are dependent on timing of exposure; embryonic ATZ exposure alters brain development and function; and embryonic ATZ exposure changes protein abundance in carcinogenesis-related pathways. After exposing embryos to 0, 0.3, 3, or 30 parts per billion (ppb) ATZ, we monitored the expression of cytochrome P450 family 17 subfamily A member 1 (cyp17a1), glyoxalase I (glo1), ring finger protein 14 (rnf14), salt inducible kinase 2 (sik2), tetratricopeptide domain 3 (ttc3), and tumor protein D52 like 1 (tpd52l1) at multiple embryonic time points to determine normal expression and if ATZ exposure altered expression. Only cyp17a1 had normal dynamic expression, but ttc3 and tpd52l1 had ATZ-related expression changes before 72 h. Larvae exposed to 0.3 ppb ATZ had increased brain length, while larvae exposed to 30 ppb ATZ were hypoactive. Proteomic analysis identified altered protein abundance in pathways related to cellular function, neurodevelopment, and genital-tract cancer. The results indicate embryonic ATZ toxicity involves interactions of multiple pathways., Significance: This is the first report of proteomic alterations following embryonic exposure to atrazine, an environmentally persistent pesticide and common water contaminant. Although the transcriptomic alterations in larval zebrafish with embryonic atrazine exposure have been reported, neither the time at which gene expression changes occur nor the resulting proteomic changes have been investigated. This study seeks to address these knowledge gaps by evaluating atrazine's effect on gene expression through multiple time points during embryogenesis, and correlating changes in gene expression to pathological alterations in brain length and functional changes in behavior. Finally, pathway analysis of the proteomic alterations identifies connections between the molecular changes and functional outcomes associated with embryonic atrazine exposure., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Short-term reproduction assays were conducted with fathead minnow (Pimephales promelas) and Japanese medaka (Oryzias latipes) to evaluate responses from atrazine exposure at environmentally relevant concentrations and above. Breeding groups of fish with multiple males and females were exposed to atrazine under flow-through conditions. Fathead minnows were exposed to mean measured concentrations of 1.0, 10, 26, 52, and 105 μg atrazine/L for 28 days. Medaka were exposed to mean measured concentrations of 9.4, 48, 74, 97, and 244 μg atrazine/L for 28 or 29 days. Fish were evaluated for survival, fecundity, fertility, total length, wet weight, secondary sex characteristics, gonadosomatic index (GSI) (P. promelas only), plasma or hepatic vitellogenin (VTG), and histopathology of gonads. General observations of health and behaviour were also conducted. There were no statistically significant effects (i.e., p < 0.05) of atrazine on survival, size, reproduction, behaviour, GSI, VTG, or secondary sex characteristics in either species at any exposure level. In fathead minnows, there were no histopathological findings associated with atrazine exposure in male fish, but there was an increased proportion of Stage 4.0 ovaries accompanied by an increase in proportion of Grade 3 post-ovulatory follicles in females of the 105 μg/L treatment group. Without a concomitant increase in oocyte atresia, neither of these findings are considered adverse for the health of the fish. In medaka, there were no significant effects of atrazine exposure on histopathology in either sex. These data support current weight-of-evidence assessments that atrazine does not cause direct adverse effects on fish reproduction at environmentally realistic concentrations., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Atrazine induces the production of reactive oxygen species (ROS), which are detoxified by enzymatic and nonenzymatic mechanisms in plants. Arbuscular mycorrhizal fungi improve on this first level of plant resistance to environmental stresses through the antioxidant defense system, but the way in which nonenzymatic antioxidants relate to atrazine in arbuscular mycorrhizal roots is not well-known. In this study, a symbiotic relationship between Funneliformis mosseae and Medicago sativa L. roots was established successfully. Then, a non-targeted metabolite analysis which was hypothesis-free concerning particular metabolites was used to provide a comprehensive metabolic fingerprint and to subsequently identify, quantify, and finally find different nonenzymatic antioxidants in mycorrhizal and non-mycorrhizal roots following atrazine addition. Tocotrienol, (iso)flavonoids, and their derivate concentrations in F. mosseae-M. sativa mycorrhizal roots were significantly higher than in non-mycorrhizal roots. The majority of (iso)flavonoids and their derivates increased significantly via methylation or glycosylation, but dehydroascorbic acid in its oxidized form decreased significantly in mycorrhizal roots. In general, F. mosseae colonization results in significantly greater nonenzymatic antioxidant tocotrienol and (iso)flavonoids derivate concentrations in M. sativa roots, which may be associated with resistance to atrazine.

Atrazine (ATR) is a widely used herbicide that has been implicated as a neurotoxicant. Recent experimental evidence has implicated that ATR exposure also appears to have adverse effects on the hippocampus, which is a critical region for learning and memory. The aim of the present study was to investigate the effects of ATR toxicity on the hippocampus of developing rats. Postnatal day (PND) 28 male Sprague⁻Dawley (SD) rats received ATR by oral gavage at 10 or 100 mg/kg bodyweight (BW) for 30 consecutive days and were sacrificed at PND 90. Behavioral test results indicated that spatial learning and memory were affected by ATR treatment. Electron microscopy analysis showed that the ultrastructures of the hippocampus were altered in the ATR-treated groups, as compared to the control group. Additionally, ATR treatment impacted dopamine and D1 dopamine receptor (D1DR) contents through different mechanisms. Reduced mRNA and protein expression levels of factors involved in the cAMP-dependent signaling pathway were also detected. These results indicate that the developmental exposure of rats to ATR can damage the hippocampus and spatial memory, which might be related to the downregulation of expression levels of the D1DR and its downstream signaling pathway.

We developed a high-resolution expression microarray based on 2456 unique transcripts from a cDNA library of the aquatic midge (Chironomus tentans). By using the microarray, we detected that 146, 434 and 243 genes were differentially expressed after C. tentans larvae were exposed to chlorpyrifos (organophosphate insecticide) at 0.1 and 0.5 μg/L, and atrazine (triazine herbicide) at 1000 μg/L, respectively, for 48 h. The number of differentially expressed genes in the larvae exposed to chlorpyrifos at 0.5 μg/L was three times of that in the larvae exposed to chlorpyrifos at 0.1 μg/L. Among the differentially expressed genes in response to chlorpyrifos exposures, 76 genes showed significant Blast hits, and among them 42 were in common between the chlorpyrifos and atrazine exposures. In 19 differentially expressed xenobiotic detoxification genes, 16 were significantly up-regulated in the larvae exposed to chlorpyrifos and/or atrazine. Two cytochrome P450 genes (CtCYP6EV1 and CtCYP4DG2) were specifically up-regulated by chlorpyrifos, whereas three cytochrome P450 genes (CtCYP4DG1, CtCYP6EX3 and CtCYP6EV3) were specifically up-regulated by atrazine. Our results showed that chlorpyrifos exposures even at low concentrations can lead to significant changes in gene expression. The significant transcriptional responses are likely attributed to larval intoxication by the insecticide. These results not only support our previous studies in which candidate gene approaches were used, but also can potentially help develop specific molecular markers for monitoring pesticide exposures in non-target organisms in aquatic systems., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Schistosomiasis is a snail-borne parasitic disease that ranks among the most important water-based diseases of humans in developing countries. Increased prevalence and spread of human schistosomiasis to non-endemic areas has been consistently linked with water resource management related to agricultural expansion. However, the role of agrochemical pollution in human schistosome transmission remains unexplored, despite strong evidence of agrochemicals increasing snail-borne diseases of wildlife and a projected 2- to 5-fold increase in global agrochemical use by 2050. Using a field mesocosm experiment, we show that environmentally relevant concentrations of fertilizer, a herbicide, and an insecticide, individually and as mixtures, increase densities of schistosome-infected snails by increasing the algae snails eat and decreasing densities of snail predators. Epidemiological models indicate that these agrochemical effects can increase transmission of schistosomes. Identifying agricultural practices or agrochemicals that minimize disease risk will be critical to meeting growing food demands while improving human wellbeing.

Background: Palmer amaranth (Amaranthus palmeri) is an economically troublesome, aggressive and damaging weed that has evolved resistance to six herbicide modes of action including photosystem II (PS II) inhibitors such as atrazine. The objective of this study was to investigate the mechanism and inheritance of atrazine resistance in Palmer amaranth., Results: A population of Palmer amaranth from Kansas (KSR) had a high level (160 - 198-fold more; SE ±21 - 26) of resistance to atrazine compared to the two known susceptible populations MSS and KSS, from Mississippi and Kansas, respectively. Sequence analysis of the chloroplastic psbA gene did not reveal any known mutations conferring resistance to PS II inhibitors, including the most common Ser264Gly substitution for triazine resistance. However, the KSR plants rapidly conjugated atrazine at least 24 times faster than MSS via glutathione S-transferase (GST) activity. Furthermore, genetic analyses of progeny generated from reciprocal crosses of KSR and MSS demonstrate that atrazine resistance in Palmer amaranth is a nuclear trait., Conclusion: Although triazine resistance in Palmer amaranth was reported more than 20 years ago in the USA, this is the first report elucidating the underlying mechanism of resistance to atrazine. The non-target-site based metabolic resistance to atrazine mediated by GST activity may predispose the Palmer amaranth populations to have resistance to other herbicide families, and the nuclear inheritance of the trait in this dioecious species further exacerbates the propensity for its rapid spread. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

The effect of atrazine and linuron, the popular and widely used chlorinated herbicides, on both myometrial contractions and secretory functions of bovine uterus and ovaries in vitro, was investigated. The pesticides inhibited (P<0.05) the basal and oxytocin (OT)-stimulated myometrial strips contractions, as well as the effect of OT on secretion of prostaglandins (PGs: PGF2α and PGE2) from endometrium. But only linuron inhibits the effect of OT on myometrial contractions. Neither of herbicides affected PGs secretion from myometrium and PGF2α from endometrium. Only the lowest dose of both tested compounds decreased PGE2 secretion from endometrium. The pesticides increased (P<0.05) the OT secretion from granulosa. However, only linuron stimulated (P<0.05) the OT secretion from the luteal cells, and it increased (P<0.05) the expression of mRNA for the OT precursor. Both compounds stimulated (P<0.05) the secretion of testosterone and atrazine increased (P<0.05) also the secretion of estradiol from the granulosa cells. While atrazine and linuron reduced (P<0.05) the progesterone secretion from the luteal cells. The data show that atrazine and linuron altered the secretory functions of ovarian cells and inhibited the myometrial contractions in vitro., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The present study sought to assess the response of Rhamdia quelen juveniles (6-8 cm total body length) to exposure to different concentrations of three herbicides: Roundup ® Original (18, 36, 72, and 144 μg/L), Primoleo ® (2.5, 5, 10, and 15 μg/L), and Facet ® (1.75, 3.5, 7, and 14 μg/L). Total protein (TP), glycogen (GG), total lipids (TL), triacylglycerols (TAG), lipid peroxidation (TBARS), and activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) in gills, liver, kidneys, and muscle were measured by spectrophotometry. Roundup ® (glyphosate) reduced the TP, GG, and TL in gills and TL in liver and kidney and increased TP in liver and increased GG in muscle. In contrast to Primoleo ® (atrazine), all tissues stored TAG and consumed LT, besides the gills also reduced PT. There was still an increase in GG in the kidneys and muscle. Facet ® (quinclorac) induced changes mainly in the liver (increased TP, TL, and TAG content) and muscle (increased GG, TL, and TAG depletion). Gill tissue exhibited TP depletion alone, and kidney tissue metabolism was unchanged. This fish species appears capable of modulating its enzymes to the point where it sustains no oxidative damage as a result of exposure to the herbicides glyphosate (possibly due to increased CAT activity), atrazine (despite no changes in SOD or CAT activity), and quinclorac (with increased lipid peroxidation, particularly in gill, kidney, and muscle tissue, despite elevated SOD activity). Although it is not considered a target species, R. quelen suffers harmful effects from interaction with these herbicides., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objective: To elucidate the effects of atrazine( ATZ) on apoptosis in cultured rat Sertoli cells., Methods: The Sertoli cells isolated from male SD rats( 18-21day) were used to set up Sertoli cells primary culture model in vitro, and these cells were treated with 0, 10, 50, 100 and 200 μmol / L ATZ for 24 h. Cell viability was assessed by MTT assay, and the vimentin expression of Sertoli cells was analyzed by immunofluorescence assay, and apoptosis were evaluated by flow cytometry. Real timePCR was used to analyze the mRNA levels of Fas L, caspase-3 and caspase-8, and the protein contents of Fas L, caspase-3 and caspase-8 were determined by ELISA., Results: The viability of Sertoli cells of 50, 100 and 200 μmol / L ATZ groups were 82. 47%, 75. 23% and 53. 76%. Compared with the control group, the viability of Sertoli cells decreased by 17. 53%( P < 0. 05), 24. 77%( P < 0. 05) and 46. 24%( P < 0. 01), respectively. The apoptosis rates of Sertoli cell of 50, 100 and 200 μmol / L ATZ groups were 8. 53%, 13. 07% and 14. 45%. Compared with the control group, the apoptosisrates of Sertoli cells were increased by 86. 86%( P < 0. 01), 186. 13%( P < 0. 01), 216. 06%( P < 0. 01), respectively. the mRNA and protein expressions of Fas L, caspase-3 and caspase-8 were significantly increased at 50, 100 and 200 μmol / L ATZ groups( P < 0. 05, P < 0. 01). The Vimentin expression of Sertoli cells was inhibited in treated groups., Conclusion: ATZ can affect the viability of Sertoli cells, and induce apoptosis of Sertoli cells through a Fas L dependent pathway.