Agent orange exposure and prostate cancer risk in the million veteran program.

BACKGROUND: The US government considers veterans to have been exposed to Agent Orange if they served in Vietnam while the carcinogen was in use, and these veterans are often deemed at high risk of prostate cancer (PCa). Here, we assess whether presumed Agent Orange exposure is independently associated with increased risk of any metastatic or fatal PCa in a diverse Veteran cohort still alive in the modern era (at least 2011), when accounting for race/ethnicity, family history, and genetic risk. PATIENTS AND METHODS: Participants in the Million Veteran Program (MVP; enrollment began in 2011) who were on active duty during the Vietnam War era (August 1964-April 1975) were included (n = 301,470). Agent Orange exposure was determined using the US government definition. Genetic risk was assessed via a validated polygenic hazard score. Associations with age at diagnosis of any PCa, metastatic PCa, and death from PCa were assessed via Cox proportional hazards models. RESULTS AND INTERPRETATION: On univariable analysis, exposure to Agent Orange was not associated with increased PCa (hazard ratio [HR]: 1.02, 95% confidence interval [CI]: 1.00-1.04, p = 0.06), metastatic PCa (HR: 0.98, 95% CI: 0.91-1.05, p = 0.55), or fatal PCa (HR: 0.94, 95% CI: 0.79-1.09, p = 0.41). When accounting for race/ethnicity and family history, Agent Orange exposure was independently associated with slightly increased risk of PCa (HR: 1.06, 95% CI: 1.04-1.09, <10-6) but not with metastatic PCa (HR: 1.07, 95% CI: 0.98-1.15, p = 0.10) or PCa death (HR: 1.02, 95% CI: 0.83-1.23, p = 0.09). Similar results were found when accounting for genetic risk. Agent Orange exposure history may not improve modern PCa risk stratification.

Assessment of the effects of 2,4-D-dichlorophenoxyacetic acid-based herbicide exposure in eggs and embryos of golden apple snails using OCT.

This study aimed to evaluate the effects of herbicide 2, 4-D-dichlorophenoxy acetic acid on golden apple snail eggs and embryos. Additionally, the study assessed the applicability of optical coherence tomography (OCT), a non-invasive depth cross-sectional microscopic imaging technique, as a novel method, to the best of our knowledge, for studying morphological changes in golden apple snail eggs and embryos, in comparison to the conventional approach of using white light microscopy. The study revealed that the herbicide 2,4-D-dichlorophenoxy acetic acid affected the hatchery rate and morphological changes of the eggs and embryos. The lethal concentration (LC50), representing the concentration of a substance that is expected to cause death in half of the population being studied, of the golden apple eggs and embryos increased with longer exposure time and higher concentrations. The estimated median effective concentration (EC50), which denotes the concentration producing the desired effect in 50% of the exposed golden apple embryos, exhibited a similar trend of change as the LC50. When compared to the microscopic study, it was observed that OCT could be employed to investigate morphological changes of golden apple snail eggs and embryos, enabling evaluation of alterations in both 2D and 3D structures.

Biomonitoring of 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide: A global view.

We conducted a literature review of urinary 2,4-D in populations not associated with a herbicide application. Of the 33 studies identified, the median/mean concentrations were similar for children, adults, and pregnant women regardless of geography. Individuals with highest concentrations may have had opportunities to directly contact 2,4-D outside of an application. Most studies were conducted in populations in North America and did not examine potential sources of 2,4-D, or what factors might influence higher or lower urinary 2,4-D concentrations. In the future, prioritizing the examination of 2,4-D biomonitoring in other regions and collecting information on sources and factors influencing exposures would better our understanding of 2,4-D exposures globally. In all the studies reviewed the concentrations of urinary 2,4-D observed were orders of magnitude below the US regulatory endpoints, suggesting that people are not being exposed to 2,4-D at levels high enough to result in adverse health effects.

New insights about the toxicity of 2,4-D: Gene expression analysis reveals modulation on several subcellular responses in Chironomus riparius.

Herbicides are the main class of pesticides applied in crops and are capable of polluting the surrounding freshwater system; thus, understanding their impact on non-target species, whose mechanism of action is not described, helps to elucidate the real risks of these pollutants to the environment. 2,4-dichlorophenoxyacetic acid (2,4-D) is frequently detected in water and, due to its persistence, poses a risk to wildlife. In this way, the present work aimed to describe the implication of exposure to concentrations of 2,4-D already reported in aquatic environments in several physiological mechanisms of C. riparius at molecular and biochemical levels. To achieve this, bioassays were conducted with fourth instar larvae exposed to three concentrations of 2,4-D (0.1, 1.0, and 7.5 µg L-1). Larvae were collected after 24 and 96 h of exposure, and the expression of 42 genes, related to six subcellular mechanisms, was assessed by Real-Time PCR (RT-PCR). Besides, the activity of the enzymes catalase (CAT), glutathione S-transferase (GST), and acetylcholinesterase (AChE) was determined. The main metabolic route altered after exposure to 2,4-D was the endocrine system (mainly related to 20-hydroxyecdysone and juvenile hormone), confirming its endocrine disruptor potential. Four of the eleven stress response genes studied were down-regulated, and later exposure modulated DNA-repair genes suggesting genotoxic capacity. Moreover, only one gene from each detoxification phase was modulated at short exposure to 1.0 µg L-1. The molecular responses were not dose-dependent, and some early responses were not preserved after 96 h, indicating a transient response to the herbicide. Exposure to 2,4-D did not alter the activity of CAT, GST, and AChE enzymes. The responses described in this study reveal new mechanistic pathways of toxicity for 2,4-D in non-target organisms and highlight potential ecological consequences for chironomids in aquatic systems at the edges of agricultural fields.

A mixture of glyphosate and 2,4-D herbicides enhances the deleterious reproductive outcomes induced by Western diet in obese male mice.

The consumption of Western diet (WD) - enriched in fats and sugars - is associated with overweight, obesity and male reproductive disorders. In addition to WD intake, crops and dairy products display residues of herbicides, including glyphosate and 2,4-D that are widely applied worldwide. The concomitant exposure to WD and herbicides - mimicking contemporary scenarios - is not fully investigated. Thus, we evaluated the effects of glyphosate and 2,4-D, alone or in mixture, on WD-induced alterations in the male genital system. Male C57BL6J mice were submitted to WD (chow containing 20% lard, 0.2% cholesterol, 20% sucrose, and high sugar solution with 23.1 and 18.9 g/L of D-fructose and D-glucose) for 6 months. Concomitantly to WD, the animals received glyphosate (0.05, 5, or 50 mg/kg/day), 2,4-D (0.02, 2 or 20 mg/kg/day) or their mixture (0, 05 + 0.02, 5 + 2, or 50 + 20 mg/kg/day) by intragastrical administration (5×/week). Doses were based on Acceptable Daily Intake (ADIs) or No Observed Adverse Effect Level (NOAEL) values. Herbicide exposure did not alter the WD-induced obesity, hypercholesterolemia and hyperglycemia. WD induced sperm cell abnormalities, reduced the number, volume and area of Leydig cells, enhanced the frequency of epididymal abnormalities, decreased the proliferation in both germinal and epididymal epithelia, and reduced the number of androgen receptor (AR) positive epididymal cells. Remarkably, the herbicide mixtures promoted such WD-induced effects: increased the frequency of sperm cell and epididymal abnormalities (absence of sperm, cytoplasmic vacuoles, and clear cell hypertrophy) (5 + 2 and 50 + 20 doses); decreased Leydig cell nuclei volume and area (5 + 2 and 50 + 20 doses), reduced epididymal cell proliferation (all mixtures), and AR expression (50 + 20 dose). In addition, herbicide mixtures reduced serum testosterone levels (5 + 2 and 50 + 20 doses). Our findings indicate that the mixture of glyphosate and 2,4-D herbicides, mimicking environmentally relevant scenarios, promotes WD-induced changes in the male genital system.

Ecological Implications on Aquatic Food Webs Due to Effects of Pesticides on Invertebrate Predators in a Neotropical Region.

Predation presents specific behavioral characteristics for each species, and the interaction between prey and predator influences the structuring of the food web. Concerning insects, predation can be affected in different ways, such as exposure to chemical stressors, e.g., pesticides. Therefore, analyses were carried out of the effects of exposure to insecticide fipronil and the herbicide 2,4-D on predation, parameters of food selectivity, and the swimming behavior of two neotropical predatory aquatic insects of the families Belostomatidae (giant water bugs) and larvae of Libellulidae (dragonfly). These predatory insects were exposed for 24 h to a commercial formulation of the chlorophenoxy herbicide, 2,4-D at nominal concentrations of 200, 300, 700, and 1400 µg L-1, and to a commercial formulation of the phenylpyrazole insecticide, fipronil at nominal concentrations of 10, 70, 140, and 250 µg L-1. In a control treatment, the insects were placed in clean, unspiked water. At the end of the exposure, the maximum swimming speed of the predators was evaluated. Afterward, the predators were placed in clean water in a shared environment for 24 h with several prey species, including the cladoceran Ceriodaphnia silvestrii, larvae of the insect Chironomus sancticaroli, the amphipod Hyalella meinerti, the ostracod Strandesia trispinosa, and the oligochaete Allonais inaequalis for 24 h. After this period, the consumed prey was counted. The results reveal that predators from both families changed prey consumption compared with organisms from the control treatment, marked by a decrease after exposure to fipronil and an increase in consumption caused by 2,4-D. In addition, there were changes in the food preferences of both predators, especially when exposed to the insecticide. Exposure to fipronil decreased the swimming speed of Belostomatidae individuals, possibly due to its neurotoxic effect. Exposure to the insecticide and the herbicide altered prey intake by predators, which could negatively influence the complex prey-predator relationship and the functioning of aquatic ecosystems in contaminated areas.

Effects of glyphosate, 2,4-D, chlorantraniliprole, and imidacloprid formulations, separately and in mixtures in Caiman latirostris hatchlings.

The present study demonstrated the potential of glyphosate (GLY), 2,4-dichlorophenoxyacetic acid (2,4-D), imidacloprid (IMI) and chlorantraniliprole (CAP) separately and in mixtures to induce oxidative stress and DNA damage in Caiman latirostris hatchlings. Under controlled condition, an embryonic exposure to these pesticides was done at concentrations recommended for soybean crops. Treatments were: negative control, GLY, 2,4-D, IMI, CAP, mixture 1 (M1): GLY + 2,4-D, M2: IM I + CAP and M3: GLY + 2,4-D + IMI + CAP. At hatching, blood samples were taken for the evaluation of genotoxicity, oxidative damage to lipids and DNA, the enzymatic activity of catalase (CAT) and superoxide dismutase (SOD), and the expression level of their corresponding genes (catalase: cat and superoxide dismutase: sod). It has been shown that IMI, M2 and M3 induced a significant inhibition of CAT activity while no effect was observed on SOD. In turn, lipid peroxidation was significantly higher in individuals exposed to IMI, and to all the mixtures. Besides, genotoxicity and oxidative DNA damage were observed in all exposed groups. The results of mRNA expression showed no difference at transcription levels. In the same way, no alterations in growth parameters were recorded at hatching. Regarding to the mixtures, we observed a potentiating action of IMI on M3 in lipid peroxidation as well as independent action on oxidative DNA damage and genotoxicity parameters. Our results highlight the importance of investigating the effect of pesticides and their mixtures considering the potential consequences to caimans living in natural environments.

Modifying effects of 2,4-D and Glyphosate exposures on gut-liver-adipose tissue axis of diet-induced non-alcoholic fatty liver disease in mice.

Nonalcoholic fatty liver disease (NAFLD), which is linked to western diet (WD) intake, affects 30% of the world's population and involves the crosstalk of liver steatosis, hypertrophy/inflammation of adipose tissue and deregulation of gut microbiome. Glyphosate and 2,4-D are some of the most applied herbicides worldwide, and their roles in NAFLD have not been investigated. Thus, the present study evaluated whether glyphosate and 2,4-D, in single or mixed exposure, alter WD-induced NAFLD in a mouse model. Male C57Bl/6 mice (n = 10/group) received a fat (30% lard, 0.02% cholesterol), and sucrose-rich diet (20%) and high sugar solution (23.1 and 18.9 g/L of fructose and glucose) for 6 months. Simultaneously, animals received glyphosate (0.05 or 5 mg/kg/day), 2,4-D (0.02 or 2 mg/kg/day), or their combination (0.05 +0.02 or 5 +2 mg/kg/day) by intragastrical administration (5 ×/week). Doses were based on the Acceptable Daily Intake (ADIs) or No Observed Adverse Effect Level (NOAEL) levels. Herbicide exposures featured differential responses. WD-induced obesity, hypercholesterolemia, and hyperglycemia remained unaltered. Compared to the group receiving only WD, only the concomitant exposure to WD and 2,4-D (2 mg) enhanced the percentage of mice with moderate/severe hepatic inflammation, CD68 macrophage infiltration, and malondialdehyde levels in the liver. In line, this herbicide modulated immune response- (including Cd4, C8b, Cd28, Cxcr3, Cxcr6) and oxidative stress-related (such as Gsta1, Gsta2, Gsta4, Gstm1, Gstm2, Gstm3, Gstm4, Nqo1, Gpx2) genes in the hepatic transcriptome analysis. This exposure also enriched pro-inflammatory Deferribacteres phylum in fecal microbiome. In general, the herbicide mixtures did not feature the same effects attributed to 2,4-D isolated exposure. Our findings indicate that 2,4-D, at a dose within the toxicological limits, was able to induce disturbances in mainly at the liver and gut axes involved in NAFLD development in male mice.

Spatiotemporal variations of 2,4-dichlorophenoxy acetic acid with the role of sugarcane industry and related human health risk assessments in the Shadegan International Wetland.

2,4-Dichlorophenoxyacetic acid (2,4-D) is the most important pesticide widely used in Khuzestan province. This study aimed to determine the ecological risks assessment in fish, plant tissues, and trends in 2,4-D urinary biomarker concentration in humans of Shadegan International wetland, Iran. Sampling was carried out from three areas: freshwater, saltwater, and brackish water. The average concentration of 2,4-D at point 38, taken from the wetland's northern parts, was 15.73 µg/L. In the lower regions, it reached 326 µg/L at point 25. From points 37 to 16, it was higher than the international standard. The increasing trend of 2,4-D concentration was observed from point 36 to the middle of the wetland. The non-carcinogenic risk through ingestion and skin contact for children and adults was 3E-3 and 1.5E-2, 2.1E-3, and 8.6E-5, respectively, and through fish, consumption was 7.1E-4 for both groups. HI values were below one (< 1), indicating no health risk. HQ values in the summer were more than one (> 1), indicating the high risk to aquatic organisms and human health.

Soybean injury caused by the application of subdoses of 2,4-D or dicamba, in simulated drift.

2,4-D or dicamba can cause injuries and other deleterious effects on non-tolerant soybeans. Thus, the objective was to evaluate the potential for injury of subdoses of 2,4-D or dicamba, in drift simulation, for application in non-tolerant soybeans. Two experiments were carried out, one with 2,4-D and the other with dicamba. The treatments consisted of the application, in post-emergence of non-tolerant soybean, of subdoses 0; 1.35; 2.68; 5.37; 10.72; 21.45 and 42.9 g acid equivalent (ae) ha-1 2,4-D choline salt or dicamba diglycolamine (DGA) salt. Injury symptoms in plants, plant height and yield were evaluated, and the results were subjected to regression analysis. Polynomial fit was possible for the doses of both herbicides, with deleterious effects on soybean, with reductions in height and yield. The application of 2,4-D ≥ 10.72 g ae ha-1 was enough to cause injuries greater than 10% in plants, in simulated drift. The application of dicamba ≥1.35 g ae ha-1 was enough to cause injuries greater than 30% in plants, in simulated drift. For both herbicides, greater potential for injury and reductions in soybean yield were observed for the application of the highest doses (21.45 and 42.9 g ae ha-1).

Protective effect of Thymus munbyanus aqueous extract against 2,4-dichlorophenoxyacetic acid-induced nephrotoxicity in Wistar rats.

The aim of the present study was to examine the possible protective effects of the aqueous extract of Thymus munbyanus (TMAE) against 2,4-dichlorophenoxyacetic acid (2,4-D)-induced oxidative stress and renal injury in the kidney of male albino rats. Furthermore, TMAE was assessed to determine the phenolic content. In vitro, antioxidant activities were evaluated by DPPH radical scavenging, inhibition of ß-carotene bleaching, and reducing power. The results showed that TMAE contained high amounts of phenolics, flavonoids, and tannins. Additionally, 24 rats were randomly divided into four groups: a control group, and three groups treated with TMAE (10 mL/kg body weight), 2,4-D (5 mg/kg body weight), and 2,4-D/combined with the TMAE for 4 weeks. Treatment with 2,4-D induced kidney dysfunctions demonstrated as an increase in the potential markers of renal filtration, namely urea and creatinine, associated with a decrease in uric acid. Moreover, 2,4-D increased malondialdehyde and carbonyl protein levels. Additionally, reduced glutathione (GSH) content, as well as superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione S-transferase (GST) activities were significantly decreased. Our results demonstrated that oral TMAE supplementation in 2,4-D-treated rats improved and restored some blood parameters and alleviated the adverse cytotoxic effects of 2,4-D by increasing certain antioxidants, consequently attenuating the intensity of oxidative stress induced by 2,4-D; this was confirmed by the histological improvements observed in the kidneys. In conclusion, TMAE demonstrated potential as a natural antioxidant, effectively alleviating 2,4-D induced kidney oxidative injury.

2,4-dichlorophenoxyacetic acid induces ROS activation in NLRP3 inflammatory body-induced autophagy disorder in microglia and the protective effect of Lycium barbarum polysaccharide.

The pesticide 2,4-dichlorophenoxyacetic acid (2,4-D) exerts neurotoxic effects; however, its action mechanism remains unclear. Here, we used BV2 cells as a model and divided them into six groups: control group (serum-free medium), lipopolysaccharide (LPS) (1 µg/mL), 2,4-D (1.2 µmol/mL), Lycium barbarum polysaccharide (LBP; 300 µg/mL LBP), LPS (1 µg/mL) + LBP (300 µg/mL), and 2,4-D (1.2 µmol/mL) + LBP (300 µg/mL) with dimethyl sulfoxide as the solvent. Our results showed that 2,4-D treatment decreased superoxide dismutase and glutathione peroxidase activities and increased malondialdehyde content. The percentage of microglial activation (co-expression of ionized calcium-binding adaptor protein-1 + CD68) in the LPS and 2,4-D groups and the levels of tumor necrosis factor alpha, interleukin (IL) 1 beta, IL-6, and IL-18 in the cell supernatant were increased. The protein and mRNA levels of Nod-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein, caspase-1, IL-1ß, IL-18, and p62 increased, whereas those of LC3II/I and Beclin-1 decreased in the 2,4-D group. The protein expression and mRNA levels of NLRP3, cleaved caspase-1, IL-1ß, IL-18, and p62 decreased significantly, whereas the protein expression and mRNA levels of LC3II/I and Beclin-1 increased in small interfering RNA of NLRP3-treated BV2 cells stimulated with 2,4-D and LPS. In conclusion, 2,4-D enhanced cell migration, promoted oxidative stress, induced excessive release of mitochondrial reactive oxygen species, promoted microglial cell activation, released inflammatory factors, activated NLRP3 inflammasomes, and inhibited autophagy. Meanwhile, LBP reduced inflammation and the release of mitochondrial reactive oxygen species, inhibited NLRP3 inflammasome activation, and regulated autophagy, thereby playing a neuroprotective role.

Mechanism of 2,4-Dichlorophenoxyacetic acid-induced damage to rat testis via Fas/FasL pathway and the protective effect of Lycium barbarum polysaccharides.

The herbicide 2,4-Dichlorophenoxyacetic acid (2,4-D) is widely used to control broadleaved weeds and has been associated with male infertility. We studied the molecular mechanisms of 2,4-D induced male reproductive system damage and the protective effects of Lycium barbarum polysaccharides (LBP) using Sprague Dawley rats and TM4 cells. Treatment with 2,4-D caused architectural and functional changes in the testis, including collapsed and atrophied seminiferous tubules with reduced number of spermatozoa, scarce sperm in the epididymal duct, low levels of serum testosterone, decreased superoxide dismutase and glutathione peroxidase activity, high malondialdehyde content, and increased apoptosis in the testis and epididymis. The expression of Fas, FasL, FADD, Pro-caspase-8, Cleaved-Caspase-8, Pro-Caspase-3, and Cleaved-Caspase-3 were significantly increased in the testicular tissue of 2,4-D-treated rats. The proliferative activity of TM4 cells decreased with an increase in dose and time of 2,4-D exposure, along with enhanced Fas/Fas ligand expression and a decreased concentration of inhibin B in TM4 cell culture medium. Depletion of Fas by specific shRNA transfection reversed the effects of 2,4-D in TM4 cells, further confirming the involvement of death receptor pathway in 2,4-D-mediated apoptosis of sertoli cells. Treatment with LBP also reversed the effects of 2,4-D in testicular cells, resulting in improved cell architecture along with enhanced proliferative capacity. Moreover, in response to LBP treatment of Sertoli cells, the content of inhibin B increased, the level of reactive oxygen species and malondialdehyde decreased, the activities of superoxide dismutase and glutathione peroxidase increased, and the rate of apoptosis as well as the expression of Fas/Fas ligand signaling pathway proteins decreased.

The Longitudinal Profile of a Stream Contaminated With 2,4-D and its Effects on Non-Target Species.

Pesticides can cause harmful effects to aquatic communities, even at concentrations below the threshold limit established as guidelines for the water bodies by environmental agencies. In this research, an input of the herbicide 2,4-dichlorophenoxyacetic acid (i.e., 2,4-D) was simulated under controlled conditions in a 500-m-long reach of a first-order tropical stream in Southeastern Brazil. Two water samplings at eight stations investigated the stream longitudinal contamination profile. The ecotoxicological effects were analyzed using Eruca sativa L. seed germination assays and the acute and chronic toxicity tests with the neotropical cladoceran Ceriodaphnia silvestrii. Physicochemical parameters of water quality were evaluated to characterize the study area and quantify 2,4-D concentrations along the stream to assess pesticide retention. The 2,4-D concentration was reduced by approximately 50% downstream in the samplings, indicating that the herbicide was retained along the stream. Moreover, C. silvestrii reproduction in long-term assays decreased approximately 50% in the stations with higher concentrations of 2,4-D than the laboratory control. After contamination, E. sativa L. showed a lower average root growth (1.0 cm), statistically different from the control (2.2 cm). On the other hand, similar growth values were obtained among the background and the most downstream stations. Our study highlighted the relevance of reviewing and updating herbicide guidelines and criteria to prevent possible ecological risks.

Evaluation of myenteric neurons in the colon of rats exposed to 2,4 dichlorophenoxyacetic acid herbicide.

The assessment of the enteric nervous system provides a better understanding of the effects that contaminants can have on the health and well-being of organisms. It has been reported that 2,4-dichlorophenoxyacetic acid (2,4-D) is a highly persistent herbicide in the environment that is responsible for neurotoxic changes in different myenteric neuronal subpopulations. The current study aimed to evaluate the effects of 2,4-D on myenteric neurons in the colon of Rattus norvegicus for the first time. A dose of 2,4-D (5 mg/kg/day) was administered to the experimental group (2,4-D) for 15 days. Then, the proximal colon was collected and submitted to Giemsa and NADPH-d histochemical techniques for the disclosure of total and nitrergic neurons. The 2,4-D group presented a higher density of total neurons (p = 0.05, t-test), which together with the maintenance of nitrergic neuronal density, may be related to the increase in the expression of the neurotransmitter acetylcholine by colocalization, responsible for stimulating the intestinal smooth muscle and increasing the chances of the expulsion of the harmful content present in the lumen. Over 15 days, the neurotoxic effects of 2,4-D in the myenteric plexus influenced an increase in the general population of myenteric neurons in the colon.

Cytotoxic and genotoxic effects induced by associated commercial glyphosate and 2,4-D formulations using the Allium cepa bioassay.

Studies assessing the toxicity of glyphosate and 2,4-dichlorophenoxyacetic acid mixture are scarce. The aim of this study was to evaluate the cytotoxicity and genotoxicity of the mixture of these herbicides using Allium cepa. Roots were exposed to glyphosate (1.56 and 11.66 mg mL-1), 2,4-D (0.28 and 17.5 mg mL-1) and mixture for 24 h, based on the average concentration applied in the field and the acute reference dose (ARfD) established in Brazil. Both isolated and associated herbicides induced a significative decrease in mitotic index (MI) (P < 0.0001) in all tested concentrations. Regarding the genotoxicity results, 2,4-D and the mixture showed, at concentrations applied in the field, a significative increase of chromosomal anomalies (CA) index compared to control (P < 0.0001) and glyphosate (P = 0.024 and P = 0.0002, respectively). All tested groups from the ARfD showed a significative difference compared to the control group (P < 0.0001), as well as glyphosate and 2,4-D isolated compared to the mixture (P = 0.0005 and P < 0.0001, respectively). The most observed CA were apoptotic bodies, giant cells, and nuclear erosions. We emphasize the need for further studies assessing the toxicity of these herbicides' mixture due to the distinct effects caused in different organisms.

Searching for biomarkers of early detection of 2,4-D effects in a native tree species from the Brazilian Cerrado biome.

Biodiversity in the Brazilian Cerrado biome has been declining sharply with the continued expansion of agriculture and the excessive use of herbicides. Thus, the aim of this study was to evaluate the morphophysiological and biochemical responses in Dipteryx alata plants to various doses of the herbicide 2,4-D. Specific biomarkers that characterize the phytoindicator potential of this species were determined. Gas exchange, chlorophyll a fluorescence, photosynthetic pigments, and the activities of antioxidant enzymes and cellulase were performed after 24, 96 and/or 396 hours after 2,4-D application (HAA). The herbicide caused higher antioxidant enzymatic activity 24 HAA and damage to the photosynthetic machinery after 96 HAA. Reduction in gas exchange, chlorophyll content, and photochemical traits were observed. Increased respiratory rates, non-photochemical quenching, and carotenoid concentrations in 2,4-D-treated plants were important mechanisms in the defense against the excess energy absorbed. Furthermore, the absence of leaf symptoms suggested tolerance of D. alata to 2,4-D. Nevertheless, changes in the photosynthetic and biochemical metabolism of D. alata are useful as early indicators of herbicide contamination, especially in the absence of visual symptoms. These results are important for early monitoring of plants in conserved areas and for preventing damage to sensitive species.

Using the Matrix to bridge the epidemiology/risk assessment gap: a case study of 2,4-D.

BACKGROUND: The Matrix is designed to facilitate discussions between practitioners of risk assessment and epidemiology and, in so doing, to enhance the utility of epidemiology research for public health decision-making. The Matrix is comprised of nine fundamental "asks" of epidemiology studies, focusing on the types of information valuable to the risk assessment process. OBJECTIVE: A 2,4-dichlorophenoxyacetic acid (2,4-D) case study highlights the extent to which existing epidemiology literature includes information generally needed for risk assessments and proffers suggestions that would assist in bridging the epidemiology/risk assessment gap. METHODS: Thirty-one publications identified in the US Environmental Protection Agency 2,4-D epidemiology review were assessed. These studies focused on associations between 2,4-D exposure and non-Hodgkin lymphoma (NHL), respiratory effects, and birth outcomes. RESULTS: Many of the papers met one or more specific elements of the Matrix. However, from this case study, it is clear that some aspects of risk assessment, such as evaluating source-to-intake pathways, are generally not considered in epidemiology research. Others are incorporated, but infrequently (e.g. dose-response information, harmonization of exposure categories). We indicated where additional analyses or modifications to future study design could serve to improve the translation. DISCUSSION: Interaction with risk assessors during the study design phase and using the Matrix "asks" to guide the conversations could shape research and provide the basis for requests for funds to support these additional activities. The use of the Matrix as a foundation for communication and education across disciplines could produce more impactful and consequential epidemiology research for robust risk assessments and decision-making.

Elevated pulmonary levels of Axin2 in mice exposed to herbicide 2,4-D with or without endotoxin.

2,4-Dichlorophenoxyacetic acid (2,4-D), a member of the phenoxy family of herbicides is commonly used in agriculture for controlling broadleaf weeds but its uncontrolled and incoherent use has been linked to incidences of lung toxicity. The present study aimed to understand the molecular mechanisms behind the 2,4-D alone or in combination with endotoxin (lipopolysaccharide [LPS]) induced pulmonary toxicity. Blood and lung samples were collected from Swiss albino mice (n = 48) following chronic exposure to high (37 mg/kg; 1/10th of LD50 ) and low (18.5 mg/kg; 1/20th of LD50 ) doses of 2,4-D alone or in combination with endotoxin (80 µg/animal). Transcriptome analysis revealed Wnt Canonical signaling as one of the top dysregulated pathways in mice lung following exposure to 2,4-D with and without endotoxin (LPS) co-exposure. Global view of differentially expressed genes showed increased messenger RNA expression of Axin2 by 0.26, 2.58, 3.14, 2.59, and 2.97 folds following exposure to LPS, high dose alone or in combination with LPS and low dose alone or in combination with LPS, respectively. The microarray data were validated using quantitative polymerase chain reaction and immunohistochemistry. Furthermore, the plasma concentration of Axin2 was elevated in the high dose group as revealed by Sandwich ELISA. The data taken together suggest a role of Axin2 to activate the Canonical Wnt signaling pathway in 2,4-D and or endotoxin-induced lung damage in mice.

Impact of 2,4-D and fipronil on the tropical midge Chironomus sancticaroli (Diptera: Chironomidae).

Increased use of pesticides in conventional agriculture implies potential risks to the environment. In aquatic ecosystems, benthic organisms may be exposed to pesticides via contaminated water and sediment, leading to several potential cascading effects on the food web. The aim of this study was to assess the functional implications of environmental realistic concentrations of the herbicide 2,4-D and the insecticide fipronil (alone and in combination) to the native tropical chironomid Chironomus sancticaroli. These two pesticides are widely applied to different crops and have frequently been detected (together) in surface water bodies in Brazil and elsewhere. Commercial products containing fipronil (Regent® 800WG) and 2,4-D (DMA® 806BR) were evaluated in 8-day toxicity tests for their effects on larval survival, growth (body length and biomass), head capsule width, development, and mentum deformities. Fipronil decreased the larval survival at the highest test concentration and the effective concentrations (EC) after eight days of exposure were: EC10 = 0.48 µg L-1 (0.395-0.565), EC20 = 1.06 µg L-1 (0.607-1.513), and EC50 = 3.70 µg L-1 (1.664-5.736). All sublethal test concentrations of fipronil decreased the larval growth, causing reductions in biomass up to 72%. The two highest test concentrations of fipronil decreased the head capsule width and after exposure to 3.7 µg fipronil L-1, only half of the larvae reached the fourth instar. The incidence of deformities was increased by fipronil in a concentration dependent manner with an increase ranging from 23% to 75%. The highest test concentration of 2.4-D (426 µg L-1) decreased the head capsule width, but larval development was unaffected at all concentrations evaluated. In the mixture tests, antagonism was observed at lower fipronil concentrations and synergism at higher fipronil concentrations for growth. The incidence of deformities rose with increasing fipronil concentrations. The results showed that environmental realistic concentrations of fipronil may have serious ecological implications for C. sancticaroli populations and that a mixture with the herbicide 2,4-D can have synergistic effects, potentiating the risks to the aquatic ecosystem.