Relevant Concentrations Of Atrazine Research Articles

Exposure to toxicologically relevant atrazine concentrations impair the glycolytic function of mouse Sertoli cells through the downregulation of lactate dehydrogenase

Atrazine (ATZ), a widely used herbicide with potent endocrine-disrupting properties, has been implicated in hormonal disturbances and fertility issues. Sertoli cells (SCs) play a crucial role in providing mechanical and nutritional support of spermatogenesis. Herein, we aimed to study the effects of environmentally relevant ATZ concentrations on the nutritional support of spermatogenesis provided by SCs. For that, mouse SCs (TM4) were exposed to increasing ATZ concentrations (in μg/L: 0.3, 3, 30, 300, or 3000). After 24 h, cellular proliferation and metabolic activity were assessed. Mitochondrial activity and endogenous reactive oxygen species (ROS) production were evaluated using JC-1 and CM-H2DCFDA probes, respectively. We also analyzed protein levels of lactate dehydrogenase (LDH) using Western Blot and live cells glycolytic function through Seahorse XF Glycolysis Stress Test Kit. ATZ exposure decreased the activity of oxidoreductases in SCs, suggesting a decreased metabolic activity. Although ATZ is reported to induce oxidative stress, we did not observe alterations in mitochondrial membrane potential and ROS production across all tested concentrations. When we evaluated the glycolytic function of SCs, we observed that ATZ significantly impaired glycolysis and the glycolytic capacity at all tested concentrations. These results were supported by the decreased expression of LDH in SCs. Overall, our findings suggest that ATZ impairs the glycolytic function of SCs through LDH downregulation. Since lactate is the preferential energetic substrate for germ cells, exposure to ATZ may detrimentally impact the nutritional support crucial for spermatogenesis, hinting for a relationship between ATZ exposure and male infertility.

Ecogenotoxicity of environmentally relevant atrazine concentrations: A threat to aquatic bioindicators

Atrazine (ATZ) is a herbicide that is frequently present in surface waters and may result in damage to the health of various organisms, including humans. However, most scientific literature reports injuries caused by ATZ at high concentrations, which are not found in the environment. Therefore, the scope of this study was to investigate the impacts of realistic concentrations of ATZ found in surface waters (1, 2, 5, 10, 15 and 20 μg/L) using the bioindicators Allium cepa, Daphnia magna and zebrafish (Danio rerio). ATZ elicited a genotoxic effect in A. cepa, manifested by the induction of chromosomal aberrations, and a mutagenic effect with increased incidence of micronuclei formation, promotion of cell death and reduction in nuclear size revealed by flow cytometry analysis. D. magna exposed to 10, 15 and 20 μg/L of ATZ showed significant reduction in body size after 21 days, delayed first-brood release, decreased egg production and total offspring, as well as swimming behavioral changes. ATZ exposure promoted physiological and developmental alterations in zebrafish embryos, including an increased spontaneous movement rate, which led to premature hatching at all concentrations investigated. Increase in total body length, decrease of the yolk sac area, pericardial edema and higher heart rate were also detected in ATZ-treated zebrafish. In summary, environmentally relevant concentrations of ATZ can induce substantial alterations in the three bioindicators investigated. This study evidences the deleterious effects of ATZ on three aquatic bioindicators employing established and current techniques, and may contribute to elucidate the risks caused by this widely used herbicide even at low concentrations and short-to-medium-term exposure.

Atrazine and estradiol effects on development of Acris blanchardi (Blanchard's cricket frog) exposed in outdoor enclosures.

The herbicide atrazine has been proposed as a potential endocrine disrupting compound (EDC) for amphibians. Using atrazine concentrations below or at those typically found in surface waters (0.5, 5.0, 50 μg/L), we exposed Acris blanchardi (Blanchard's cricket frog) larvae throughout development until metamorphosis (i.e. Gosner stages 26-45). An additional 50 μg/L treatment (50s μg/L) was utilized where supplemented algae was added to control for indirect atrazine effects from reduced food sources. In addition to atrazine, experimental groups also included a negative control and two positive controls, 17β-estradiol (E2) at 2.3 and 25 μg/L. At 60 days post-metamorphosis, A. blanchardi metamorphs were euthanized for analysis of gross and histopathological development. Atrazine did not significantly influence mortality (mean recovery of 54% across treatments), sex ratio, body mass (BM), snout-vent length (SVL), gonad size, nor gonad development of A. blanchardi. Females exposed to 50sμg/L atrazine had 29% less mass, were 10% shorter, and had a 29% lower mean ovary area (mm2 ) as compared to negative controls, suggesting algae enrichment had a significant negative effect. Males exposed to estradiol (25 μg/L) showed an increased level of oviduct development. Ovary area was also significantly influenced by estradiol treatment at 2.3 and 25 μg/L. Overall, estradiol had much less effect than predicted based on other model species (e.g. Xenopus laevis). Development of A. blanchardi, overall, was not affected by long-term exposure to environmentally relevant concentrations of atrazine. However, this species also was largely insensitive to exogenous estradiol in this test system. © 2022 Society of Chemical Industry.

Effects of environmentally relevant concentrations of atrazine and glyphosate herbicides, isolated and in mixture, on two generation of the freshwater microcrustacean Daphnia magna.

The herbicides atrazine and glyphosate are used worldwide and their excessive usage results in the frequent presence of these pesticides in environmental compartments. We evaluated the effects of environmentally relevant concentrations of analytical standards and commercial formulations of atrazine (2 µg L-1) and glyphosate (65 µg L-1), isolated and in mixture (2 + 65 µg L-1) on the microcrustacean Daphnia magna. Through chronic exposure (21 days) of two generations, we observed effects on survival, reproductive capacity and responses of the antioxidant defense system (catalase) and biotransformation system (glutathione S-transferase). The survival of organisms was affected in the second generation (F1) with a mortality of 17% in the mixture of commercial formulations treatments. In the evaluation of the first generation (F0) we observed only effects on sexual maturation of organisms, while in the F1, changes were observed in all parameters evaluated. A statistical difference (p < 0.05) was also observed between the analytical standards and the commercial formulations for all parameters evaluated, indicating that other components present in the formulations can change the toxicity of products. We suggest that atrazine can modulate toxicity when mixed with glyphosate, as the standard analytical atrazine and mixture of analytical standards results were similar in most parameters. Given the difficulty in estimating effects of mixtures and considering that various stressors are found in the environment, our results support the need to carry out long-term studies and, above all, to verify what are the impacts across generations, so that the toxicity of products is not underestimated. Graphical abstract.

Effects of subchronic exposure to environmentally relevant concentrations of the herbicide atrazine in the Neotropical fish Astyanax altiparanae

Atrazine (ATZ) is among the most widely used herbicides in the world, and yet it has a potential to contaminate aquatic environments due to pesticide leaching from agricultural areas. In the Neotropical region, studies about the effects of this herbicide in native aquatic wildlife is scarce.Our study aimed at investigating the effects of a 30-day exposure to a commercial atrazine formulation on oxidative stress parameters, histopathology in testis and liver, and hormone levels in males and female of yellow-tailed tetra fish (Astyanax altiparanae). Adults were exposed to low but environmentally relevant concentrations of atrazine as follows: 0 (CTL-control), 0.5 (ATZ0.5), 1 (ATZ1), 2 (ATZ2) and 10 (ATZ10) μg/L. Our results showed decreased GST activity in gills in all groups of exposed animals and increased CAT activity in gills from the ATZ10 group. In the liver, there was an increase in lipid peroxidation in fish from ATZ1 and ATZ2 groups. Histological analysis of the liver showed increased percentage of sinusoid capillaries in ATZ2 fish, increased vascular congestion in ATZ1 and increased leukocyte infiltration in the ATZ10 group. Hepatocyte diameter analysis revealed a decrease in cell size in all groups exposed to ATZ, and a decrease in hepatocyte nucleus diameter in ATZ1, ATZ2 and ATZ10 groups. Endocrine parameters did not show significant changes following ATZ exposure, although an increase of triiodothyronine/thyroxine (T3/T4) ratio was observed in ATZ2 fish. Our results provide evidence that even low, environmentally relevant concentrations of ATZ produced oxidative damage and histological alterations in adult yellow-tailed tetra.

Effects of Sub-Lethal Atrazine Concentrations on Embryogenesis, Larval Survival and Growth of African Catfish, Clarias gariepinus (Burchell, 1822)

The toxicity of environmentally relevant concentrations of atrazine on exposed fertilized eggs of Clarias gariepinus was evaluated in a 96hour static non-renewal bioassay. After exposure to atrazine concentrations of 0 (control), 4, 8, 12, and 16 μgL1 in water, first mitotic cleavage in the control occurred at 40 minutes after fertilization. The first cleavage did not occur in the 8 μgL-1 treatment group until 60 minutes after fertilization. Similar dose depended delayed cleavage was observed across atrazine exposed eggs. The first larvae emerged 22 hours from the time of fertilization at 27.00C in both control and atrazine treatment groups. However, hatching rates significantly differed between the control and atrazine exposed eggs (P&lt;0.05) with 73.3% hatching rate in the control and 3.3% among the exposed eggs treated with 16μgL-1 of atrazine. Among the morphological deformities observed in the larvae of atrazine exposed eggs, kyphosis occurred more frequently with 60% of emerged larvae in the 16 μgL-1 atrazine treated group exhibiting the deformity. Morphometry showed significantly reduced head length, body length, body weight, and yolk diameter in 16 μgL-1 atrazine exposed eggs compared to the control. These results indicate that atrazine disrupted embryonic process and delayed hatching with severe larvae deformities.

The effects of atrazine on the microbiome of the eastern oyster: Crassostrea virginica

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.

Effect of life stage and pesticide exposure on the gut microbiota of Aedes albopictus and Culex pipiens L

Pesticides commonly contaminate the aquatic environments inhabited by mosquito juveniles. However, their role in shaping the mosquito microbiota is not well understood. We hypothesized that environmentally relevant concentrations of atrazine, permethrin and malathion will mediate a shift in the mosquito gut bacterial community structure due to their toxic effect on the aquatic bacterial communities, and reduce mosquito gut bacterial diversity by enriching pesticide-degrading bacterial communities over susceptible taxa. Illumina MiSeq sequencing of the V3-V4 hypervariable regions of the 16 S rRNA gene was used to characterize the microbial communities of larval and adult stages of the two mosquito species and the water samples from microcosms treated with each of the pesticides, separately. Bacterial community composition differed by sample type (larval stage vs. adult stage) and water sampling date (day 3 vs. day 7), but not by pesticide treatment. In larval stages, bacterial OTU richness was highest in samples exposed to malathion, intermediate in permethrin, and lowest in controls. Bacterial richness was significantly higher in larval stages compared to adult stages for all treatments. This study provides a primer for future studies evaluating mosquito microbial responses to exposures to chemical pesticides and the possible implications for mosquito ecology.

Embryotoxicity of atrazine and its degradation products to early life stages of zebrafish (Danio rerio)

Triazine herbicide atrazine is considered to be moderately toxic to various aquatic animals. The aim of our study was to evaluate the acute embryotoxicity of atrazine and its two degradation products, desisopropylatrazine and desethylatrazine, and their mixture to the early life stages of zebrafish (Danio rerio) by means of a modified method of the Fish Embryo Acute Toxicity (FET) Test – OECD guideline 236. Toxic effects were studied by the evaluation of lethal endpoints and development of disorders. Furthermore, sublethal endpoints such as hatching rate, formation of somites, development of eyes, spontaneous movement, heartbeat, blood circulation, pigmentation and occurrence of edema at 24, 48, 72 and 96 h post fertilization were assessed. Newly fertilized eggs were exposed to various concentrations of atrazine, desisopropylatrazine and desethylatrazine, and their combination 0.3, 30, 100, 300, 1000, 3,000 and 10,000 μg/l, which represent environmentally relevant levels of these pollutants in surface waters and multiples of these concentrations to find out if the toxic effect depends on dose. Single substances and their combination were not associated with a negative effect on mortality. Rare malformations were observed during these embryonal toxicity tests. Only pericardial edema was recorded during the monitored observation. A significant increase in the occurrence of pericardial edema between the control 0% and the experimental group 17.6 %) was found only in the group exposed to the highest concentration of a triazine herbicide combination (10,000 μg/l) at 72 and 96 h post fertilization. Obtained results indicate that especially higher not environmentally relevant concentrations of atrazine, its metabolites or their combination present a potencial risk of embryotoxicity for zebrafish.

Cardiotoxicity in African clawed frog (Xenopus laevis) sub-chronically exposed to environmentally relevant atrazine concentrations: Implications for species survival

The toxic effects of different atrazine concentrations on tadpoles and adult male African clawed frogs (Xenopus laevis) were assessed in a controlled laboratory environment following 90 days’ exposure. The aim was to elucidate the danger of atrazine exposure on the cardiac tissue relative to its critical function of rhythmic contractility, fundamental for optimal blood circulation and homeostasis.Tadpoles and adult frogs were exposed to 0 μg/L (control), 0.01 μg L−1, 200 μg L−1 and 500 μg L−1 concentrations of atrazine for 90 days. Mortality was concenration-dependent and significantly increased in juvenile group (77%, 43%, 23% and 0 respectively for 500 μg L−1, 200 μg L−1, 0.01 μg L−1, and control group). While the mean juvenile heart area decreased concentration-dependently, adult frog mean heart area significantly increased in the 200 μg L−1 group only and mean heart weight change was variable across all exposure levels.Light microscopy of hematoxylin and eosin (H&E) and Mallory-Heidenhain rapid one-step staining techniques on cardiac tissue sections of the juvenile and adult frogs revealed shrinkage of cardiac muscle cells into thin wavy myocytes. Additionally, disorganized branching of muscle fibres with reduced striations were observed in 0.01 μg L−1 and 200 μg L−1 but hypertrophied myocytes, thickened intensely staining myofibrils in the 500 μg L−1 group in juvenile and adult frogs. Significant increase in the mean percentage area of connective tissue in all the treated groups (p < 0.036) were also recorded. Immunohistochemistry analysis showed decreased eNOS localization in cardiac tissue in 200 μg L−1 and 500 μg L−1 of both juvenile and adult group, suggestive of decreased cardiac contractility due to atrazine exposure.The results indicate that environmentally relevant atrazine concentrations cause significant mortality in tadpoles while concentrations ≥200 μg L−1 adversely affect cardiac muscle morphology and may induce functional perturbations in cardiac tissue contractility and consequent dysfunction which generally may have an adverse impact on their survival and longevity.

Acute in vitro exposure to environmentally relevant atrazine levels perturbs bovine preimplantation embryo metabolism and cell number

Atrazine is a widely used herbicide known to negatively alter endocrine systems and perturb metabolism. Preimplantation exposure to pesticides may adversely affect long-term health, however few studies examine the effect of environmental levels and whether specific periods of development are particularly sensitive. In this study, the effect of acute, preimplantation atrazine exposure (days 3.5–7.5 post-fertilization) at levels detected and deemed safe in drinking water (0.02 and 20 μg/L respectively) on in vitro bovine embryo development, quality, metabolism, and gene expression was investigated. Atrazine exposure had no effect on development or quality, but significantly reduced blastocyst total cell numbers, attributable to a decrease in trophectoderm cells. Notably, atrazine (20 μg/L) markedly increased carbohydrate metabolism. Therefore, short-term exposure to environmentally relevant atrazine concentrations perturbs bovine preimplantation embryo metabolism and cell number, highlighting a potential mechanism by which atrazine can mediate embryo viability and health.

Assessment of atrazine decontamination by epiphytic root bacteria isolated from emergent hydrophytes

Aim of the study was to identify atrazine remediating bacteria that can potentially succeed in situ where they encounter varied environmental conditions. Three epiphytic root bacteria, genus Pseudomonas and Arthrobacter, were isolated from rhizoplanes of hydrophytes Acorus calamus, Typha latifolia, and Phragmites karka. Potential of these strains to decontaminate environmentally relevant concentrations of atrazine was determined in liquid atrazine medium (LAM) and Luria-Bertani (LB) medium at varying pH and temperature. There was an increase in decontamination by the strains with time upon exposure to 2.5 to 10 mg l−1 atrazine over a period of 15 days, notably, in both minimal and nutrient-rich media. Growth in terms of O.D.600 and biomass determined during the same period also showed a corresponding surge. Pseudomonas sp. strain AACB mitigated atrazine in a wide range of pH (5 to 8). Pseudomonas sp. strains AACB and TTLB decontaminated > 62% atrazine at 10 °C. All the strains exhibited plant growth–promoting traits in vitro, reported for the first time in the presence of atrazine. Strain AACB exhibits the novel trait of atrazine decontamination under harsh environmental conditions mimicked in lab. Strains isolated in the present study promise success in in situ remediation. Bioreactors and water treatment plants can be designed comprising the hydrophytes and the strains inoculated into their rhizospheres to improve efficacy of the treatment. They can be used to study plant-bacterium mutualistic symbiosis or other interactions occurring during atrazine mitigation.

The Effects of an Herbicide and Antibiotic Mixture on Aquatic Primary Producers and Grazers.

Widespread use of agrochemicals increases their likelihood of entering aquatic systems in mixture. Despite different modes of action, atrazine (herbicide) and tetracycline (antibiotic) adversely affect non-target photosynthetic organisms individually, but the effects of simultaneous exposure to both contaminants are untested. We created microcosms containing microalgae (Chlorella sp.), floating macrophytes (Lemna minor), and a zooplankton grazer (Daphnia magna). Microcosms were exposed to environmentally relevant concentrations of atrazine and tetracycline, alone and together, for 10days. Atrazine decreased Chlorella sp. abundance, but not enough to reduce food availability to D. magna whose reproduction and mortality were unaffected. In contrast, tetracycline and atrazine appeared to have additive effects on L. minor abundance and growth inhibition. These additive adverse effects suggest increased potential for L. minor population decline over the long term, and potential for altered species interactions in aquatic systems receiving agricultural runoff.

Effects of environmentally relevant sub-chronic atrazine concentrations on African clawed frog (Xenopus laevis) survival, growth and male gonad development

Sub-chronic toxicity of environmentally relevant atrazine concentrations on exposed tadpoles and adult male African clawed frogs (Xenopus laevis) was evaluated in a quality controlled laboratory for 90 days. The aim of this study was to determine the effects of atrazine on the survival, growth and gonad development of African clawed frogs. After exposure of tadpoles to atrazine concentrations of 0 (control), 0.01, 200 and 500 μg L−1 in water, mortality rates of 0, 0, 3.3 and 70% respectively were recorded for the 90 day exposure period. Morphometry showed significantly reduced tadpole mass in the 500 μg L−1 atrazine exposed tadpoles (p < 0.05). Light microscopy on testes of adult frogs exposed to the same atrazine concentrations using hematoxylin and eosin (H&E) and Van Gieson staining techniques revealed gonadal atrophy, disruption of germ cell lines, seminiferous tubule structure damage and formation of extensive connective tissue around seminiferous tubules of frogs exposed to 200 μg L−1 and 500 μg L−1 atrazine concentrations. Ultrastructural analysis of the cellular organelles using transmission electron microscopy (TEM) revealed significant amounts of damaged mitochondria in testosterone producing Leydig cells as well as Sertoli cells. Biochemical analysis revealed reduced serum testosterone levels in adult frogs at all exposure levels as well as presence of six atrazine metabolites in frog serum and liver. The results indicate that atrazine concentrations greater than the calculated LC50 of 343.7 μg L−1 cause significant mortality in tadpoles, while concentrations ≥200 μg L−1 adversely affect reproductive health of adult frogs and development of tadpoles sub-chronically exposed to atrazine.

Atrazine feminizes sex ratio in Blanchard's cricket frogs (Acris blanchardi) at concentrations as low as 0.1 μg/L.

We exposed Blanchard's cricket frogs (Acris blanchardi) to ecologically relevant concentrations (0, 0.1, 1, and 10 μg/L) of a commercial formulation of atrazine throughout the larval period to determine effects on survival, somatic growth and development (time to metamorphosis and mass at metamorphosis), and gonadal development (sex ratio at metamorphosis and the prevalence of testicular ova in phenotypic males). We tested the following hypotheses: 1) atrazine feminizes the sex ratio, 2) atrazine increases the proportion of phenotypic males with testicular ova, and 3) atrazine differentially affects somatic growth (mass at metamorphosis) and development (time to metamorphosis) for males and females. Although the control sex ratio was male-biased, exposure to 0.1 and 10 μg/L atrazine feminized sex ratios, because these treatments produced 51 and 55% fewer males than the control, respectively. We did not observe testicular ova. Atrazine did not impact survival or metamorphosis, and we did not detect sexually dimorphic impacts on time to metamorphosis or mass at metamorphosis. However, males metamorphosed 2.3 d later than females, regardless of treatment. Sex biases in timing of metamorphosis are underexplored in anurans, but if prevalent, could have important implications for theory surrounding the impact of environmental factors on metamorphosis. Our data suggest that cricket frog sex ratios are sensitive to environmentally relevant concentrations of atrazine and that feminization in the field is likely. Environ Toxicol Chem 2018;37:427-435. © 2017 SETAC.

Short-term atrazine exposure at breeding has no impact on Blanchard's cricket frog (Acris blanchardi) reproductive success.

Studies of endocrine-disrupting contaminants have focused on early-life exposures, but later exposures could impact fitness. We exposed adult frogs (Acris blanchardi) at reproduction to ecologically relevant atrazine concentrations (0, 1, or 10 µg/L) in outdoor arenas. We measured likelihood of breeding and number of resulting tadpoles. Atrazine impacted neither the probability of breeding nor the number of tadpoles produced, suggesting anuran reproductive success may not be impacted by short-term exposure to low concentrations. Environ Toxicol Chem 2017;36:3284-3288. © 2017 SETAC.

The herbicide atrazine induces hyperactivity and compromises tadpole detection of predator chemical cues

The ability to detect chemical cues is often critical for freshwater organisms to avoid predation and find food and mates. In particular, reduced activity and avoidance of chemical cues signaling predation risk are generally adaptive behaviors that reduce prey encounter rates with predators. The present study examined the effects of the common herbicide atrazine on the ability of Cuban tree frog (Osteopilus septentrionalis) tadpoles to detect and respond to chemical cues from larval dragonfly (Libellulidae sp.) predators. Tadpoles exposed to an estimated environmental concentration of atrazine (calculated using US Environmental Protection Agency software; measured concentration, 178 μg/L) were significantly hyperactive relative to those exposed to solvent control. In addition, control tadpoles significantly avoided predator chemical cues, but tadpoles exposed to atrazine did not. These results are consistent with previous studies that have demonstrated that ecologically relevant concentrations of atrazine can induce hyperactivity and impair the olfactory abilities of other freshwater vertebrates. The authors call for additional studies examining the role of chemical contaminants in disrupting chemical communication and the quantification of subsequent impacts on the fitness and population dynamics of wildlife. Environ Toxicol Chem 2016;35:2239-2244. © 2016 SETAC.

The relationship between study sponsorship, risks of bias, and research outcomes in atrazine exposure studies conducted in non-human animals: Systematic review and meta-analysis

BackgroundA critical component of systematic review methodology is the assessment of the risks of bias of studies that are included in the review. There is controversy about whether funding source should be included in a risk of bias assessment of animal toxicology studies. ObjectiveTo determine whether industry research sponsorship is associated with methodological biases, the results, or conclusions of animal studies examining the effect of exposure to atrazine on reproductive or developmental outcomes. MethodsWe searched multiple electronic databases and the reference lists of relevant articles to identify original research studies examining the effect of any dose of atrazine exposure at any life stage on reproduction or development in non-human animals. We compared methodological risks of bias, the conclusions of the studies, the statistical significance of the findings, and the magnitude of effect estimates between industry sponsored and non-industry sponsored studies. ResultsFifty-one studies met the inclusion criteria. There were no differences in methodological risks of bias in industry versus non-industry sponsored studies. 39 studies tested environmentally relevant concentrations of atrazine (11 industry sponsored, 24 non-industry sponsored, 4 with no funding disclosures). Non-industry sponsored studies (12/24, 50.0%) were more likely to conclude that atrazine was harmful compared to industry sponsored studies (2/11, 18.1%) (p value=0.07). A higher proportion of non-industry sponsored studies reported statistically significant harmful effects (8/24, 33.3%) compared to industry-sponsored studies (1/11; 9.1%) (p value=0.13). The association of industry sponsorship with decreased effect sizes for harm outcomes was inconclusive. ConclusionOur findings support the inclusion of research sponsorship as a risk of bias criterion in tools used to assess risks of bias in animal studies for systematic reviews. The reporting of other empirically based risk of bias criteria for animal studies, such as blinded outcome assessment, randomization, and all animals included in analyses, needs to improve to facilitate the assessment of studies for systematic reviews.

Environmentally Relevant Concentrations of Atrazine and Ametrine Induce Micronuclei Formation and Nuclear Abnormalities in Erythrocytes of Fish.

A rapid and sensitive method using liquid chromatography coupled with mass spectrometry triple quadrupole direct aqueous injection for analysis of atrazine and ametrine herbicides in surface waters was developed. According to the validation method, water samples from six different locations in the Piracicaba River were collected monthly from February 2011 to January 2012 and injected into a liquid chromatographer/dual mass spectrometer without the need for sample extraction. The method was validated and shown to be precise and accurate; limits of detection and quantification were 0.07 and 0.10µg L(-1) for atrazine and 0.09 and 0.14µg L(-1) for ametrine. During the sampling period, concentrations of atrazine ranged from 0.11 to 1.92µg L(-1) and ametrine from 0.25 to 1.44µg L(-1). After analysis of the herbicides, Danio rerio were exposed a range of concentrations found in the river water to check the induction of micronuclei and nuclear abnormalities (NAs) in erythrocytes. Concentrations of atrazine and ametrine >1.0 and 1.5µg L(-1), respectively, induced MN formation in D. rerio. Ametrine was shown to be more genotoxic to D. rerio because a greater incidence of NAs was observed compared with atrazine. Therefore, environmentally relevant concentrations of atrazine and ametrine found in the Piracicaba River are dangerous to the aquatic biota.

The effects of the herbicide atrazine on freshwater snails.

Atrazine has been shown to affect freshwater snails from the subcellular to community level. However, most studies have used different snail species, methods, endpoints, and atrazine exposure concentrations, resulting in some conflicting results and limiting our understanding. The goal of this study was to address these concerns by (1) investigating the acute and chronic effects of atrazine on four species of freshwater snails (Biomphalaria glabrata, Helisoma trivolvis, Physa acuta, and Stagnicola elodes) using the same methods, endpoints, and concentrations, and (2) summarizing the current literature pertaining to the effects of atrazine on freshwater snails. We conducted a 48 h acute toxicity test with an atrazine concentration higher than what typically occurs in aquatic environments (1000 µg/L). Additionally, we exposed snails to environmentally relevant atrazine concentrations (0, 0.3, 3, and 30 µg/L) for 28 days and assessed snail survival, growth, and reproduction. We also summarized all known literature pertaining to atrazine effects on freshwater snails. The literature summary suggests snails are often affected by environmentally relevant atrazine concentrations at the subcellular and cellular levels. These effects are typically not transitive to effects on survival, growth, or reproduction at the same concentrations. Our acute exposures corroborate the general trend of no direct effect on snail populations as atrazine did not directly affect the survival of any of the four snail species. Similarly, environmentally relevant concentrations did not significantly affect the survival, growth, or reproduction of any snail species. These results indicate that, in the absence of other possible stressors, the direct effects of environmentally relevant atrazine concentrations may not be realized at the snail population level.