Complex Mixture Of Pesticides Research Articles

Low concentrations of complex mixtures of pesticides and metabolites are toxic to common Carp brain cells (Cyprinus carpio carpio)

Pesticide use increases annually, and Brazil is the world’s largest consumer. However, unlike the European Union (EU), there is no established limit value for pesticide mixtures in drinking water, and therefore the concentration of pesticides can reach 3354 times the EU limit. Thus, determining the risk of exposure to pesticide mixtures and their main metabolites is challenging and requires the use of alternative methods. In the present study, the Common Carp Brain (CCB) cell line was used to evaluate the in vitro toxicity of relevant pesticide mixtures (glyphosate, 2,4-D, atrazine, and mancozeb) and their main metabolites after 72 h of exposure. The tested concentrations were based on the Acceptable Daily Intake (ADI) defined by Brazilian legislation. The results showed that cells exposed to lower concentrations of the pesticide mixtures and the pesticide + metabolite mixtures were affected by a decrease in cell confluence, resazurin metabolism, and wound healing capacity. The IBR index showed that lower concentrations had more severe effects, suggesting the absence of safe concentrations of these pesticide and metabolite mixtures for the CCB cell line within the tested concentration range. These findings raise concerns about the effects of exposure to these substances on animal and human health.

Occurrence and toxicological relevance of pesticides and trace metals in agricultural soils, sediments, and water of the Sogamoso River basin, Colombia

Historical pesticide use in agriculture and trace metal accumulation have long term impact on soil, sediment, and water quality. This research quantifies legacy and current-use pesticides and trace metals, assessing their occurrence and toxicological implications on a watershed scale in the Sogamoso River basin, tributary of the Magdalena River in Colombia. Organochlorine pesticides (22), organophosphates (7), and azole fungicides (5), as well as trace metals cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), nickel (Ni), lead (Pb), and zinc (Zn) were analyzed in croplands and along the river. Toxic units (TU) and hazard quotients (HQ) were calculated to assess the mixture toxicity. Organochlorines were detected in 84% of soils, 100% of sediments, and 80% of water samples. Organophosphates were found in 100% of soil and sediment samples, as well as in 70% of water samples. Azole fungicides were present in 79% of soils, 60% of sediments, and in 10% of water samples. Total pesticide concentrations ranged from 214.2 to 8497.7 μg/kg in soils, 569.6–12768.2 μg/kg in sediments, and 0.2–4.1 μg/L in water. In addition, the use of partition coefficient (Kd) and organic carbon fraction (foc) allowed the distribution analysis for most of the pesticides in sediments, suspended particulate matter (SPM), and water systems, but not for soils. Concentrations of trace metals Cu, Zn, Pb, and Zn exceeded international quality guidelines for agricultural soils in 16% of the samples. Furthermore, Cu and Zn concentrations exceeded sediment quality guidelines in 50 and 90% of the samples, respectively. These findings demonstrate the broad distribution of complex mixtures of trace metals, legacy organochlorines, and current-use pesticides across the basin, indicating that conventional agriculture is a significant source of diffuse pollution. Sustainable agricultural practices are needed to mitigate adverse impacts on ecosystems and human health.

Pesticides exposure in pregnant Argentinian women: Potential relations with the residence areas and the anthropometric neonate parameters

Intrauterine environment is the first chemical exposure scenario in life, through transplacental transfer. In this study, the aim was to determine concentrations of organochlorine pesticides (OCPs) and selected current use pesticides in the placentas of pregnant women from Argentina. Socio-demographic information, the mother's lifestyle and neonatal characteristics were also analysed and related to pesticides residue concentrations. Thus, 85 placentas were collected at birth, from an area of intensive fruit production for the international market, in Patagonia Argentina. Concentrations of 23 pesticides including, trifluralin (herbicide), the fungicides chlorothalonil and HCB, and the insecticides chlorpyrifos, HCHs, endosulfans, DDTs, chlordanes, heptachlors, drins and metoxichlor, were determined by GC-ECD and GC-MS. Results were first analysed all together and then grouped by their residential settings, in urban and rural groups. Total mean pesticide concentration was 582.6 ± 1034.4 ng/g lw, where DDTs (325.9 ± 950.3 ng/g lw) and chlorpyrifos (188.4 ± 365.4 ng/g lw) showed a high contribution. Pesticide levels found exceeded those reported in low, middle and high income countries from Europe, Asia and Africa. In general, pesticides concentrations were not associated with neonatal anthropometric parameters. When the results were analysed by residence place, significantly higher concentrations of total pesticides and chlorpyrifos (Mann Whitney test p = 0.0003 and p = 0.032, respectively) were observed in placentas collected from mothers living in rural settings compared to urban areas. Rural pregnant women presented the highest pesticide burden (5.9 μg), where DDTs and chlorpyrifos were the major constituents. These results suggested that all pregnant women are highly exposed to complex pesticide mixtures, including banned OCPs and the widely used chlorpyrifos. Based on the pesticide concentrations found, our results warn of possible health impacts from prenatal exposure through transplacental transfer. This is one of the first reports of both chlorpyrifos and chlorothalonil concentrations in placental tissue, and contributes to the knowledge of current pesticide exposure in Argentina.

An original approach for the identification of relevant pesticides mixtures at nationwide scale

Wild organisms are likely exposed to complex mixtures of pesticides owing to the large diversity of substances on the market and the broad range agricultural practices. The consequences of such exposure are still poorly understood, first because of potentially strong synergistic effects, making cocktails effects not predictable from the effects of single compounds, but also because little is known about the actual exposure of organisms to pesticide mixtures in natura.

Evaluation of biochemical, hematological, and genotoxic parameters in mice exposed to individual and combined ethoprophos and cadmium

Environmental contamination by complex mixtures of pesticides and metals is a major health problem in agriculture and industry. In real life scenarios, we are exposed to mixtures of chemicals rather than single chemicals, and therefore it is critical to assess their toxicity. The current work was conducted to assess the toxic effects of a low dose (2% median lethal dose) of ethoprophos (Etho, 0.16 mg kg−1 bw), and cadmium (Cd, 0.63 mg kg−1 bw); each alone or in combination on hematological, biochemical, and genotoxic parameters in male mice for one or four weeks. The tested toxicants resulted in a decline in body and organs weights, the most hematological indices, acetylcholine esterase activity, and the total protein content, while they significantly increased liver and kidney function parameters. Furthermore, they increased the mitotic index (MI), number of abnormal sperms, and chromosomes. In conclusion, Etho and Cd induce deleterious effects on all tested parameters in male mice which reflect more obvious impacts when both combined, particularly after 28 days of exposure. However, further research is needed to confirm toxicokinetic or toxicodynamic interactions between these two toxic compounds in the organisms.

Joint toxic effects of thiamethoxam and flusilazole on the adult worker honey bees (Apis mellifera L.)

Insect pollinators are routinely exposed to a complex mixture of many pesticides. However, traditional environmental risk assessment is only carried out based on ecotoxicological data of single substances. In this context, we aimed to explore the potential effects when worker honey bees (Apis mellifera L.) were simultaneously challenged by thiamethoxam (TMX) and flusilazole (FSZ). Results displayed that TMX possessed higher toxicity to A. mellifera (96-h LC50 value of 0.11 mg a. i. L−1) than FSZ (96-h LC50 value of 738 mg a. i. L−1). Furthermore, the mixture of TMX and FSZ exhibited an acute synergistic impact on the pollinators. Meanwhile, the activities of SOD, caspase 3, caspase 9, and PPO, as well as the expressions of six genes (abaecin, dorsal-2, defensin-2, vtg, caspase-1, and CYP6AS14) associated with oxidative stress, immune response, lifespan, cell apoptosis, and detoxification metabolism were noteworthily varied in the individual and mixture challenges than at the baseline level. These data revealed that it is imminently essential to investigate the combined toxicity of pesticides since the toxicity evaluation from individual compounds toward honey bees may underestimate the toxicity in realistic conditions. Overall, the present results could help understand the potential contribution of pesticide mixtures to the decline of bee populations.

Dietary exposure to environmentally relevant pesticide mixtures impairs swimming performance and lipid homeostatic gene expression in Juvenile Chinook salmon at elevated water temperatures

Aquatic organisms are exposed to complex mixtures of pesticides in the environment, but traditional risk assessment approaches typically only consider individual compounds. In conjunction with exposure to pesticide mixtures, global climate change is anticipated to alter thermal regimes of waterways, leading to potential co-exposure of biota to elevated temperatures and contaminants. Furthermore, most studies utilize aqueous exposures, whereas the dietary route of exposure may be more important for fish owing to the hydrophobicity of many pesticides. Consequently, the current study aimed to determine the effects of elevated temperatures and dietary pesticide mixtures on swimming performance and lipid metabolism of juvenile Chinook salmon, Oncorhynchus tshawytscha. Fish were fed pesticide-dosed pellets at three concentrations and three temperatures (11, 14 and 17 °C) for 14 days and swimming performance (Umax) and expression of genes involved in lipid metabolism and energetics were assessed (ATP citrate lyase, fatty acid synthase, farnesoid x receptor and liver x receptor). The low-pesticide pellet treatment contained five pesticides, p,p’-DDE, bifenthrin, esfenvalerate, chlorpyrifos and fipronil at concentrations based on prey items collected from the Sacramento River (CA, USA) watershed, with the high-pesticide pellet treatment containing a six times higher dose. Temperature exacerbated effects of pesticide exposure on swimming performance, with significant reductions in Umax of 31 and 23% in the low and high-pesticide pellet groups relative to controls at 17 °C, but no significant differences in Umax among pesticide concentrations at 11 or 14 °C. At 14 °C there was a significant positive relationship between juvenile Chinook salmon pesticide body residues and expression of ATP citrate lyase and fatty acid synthase, but an inverse relationship and significant downregulation at 17 °C. These findings suggest that temperature may modulate effects of environmentally relevant pesticide exposure on salmon, and that pesticide-induced impairment of swimming performance may be exacerbated under future climate scenarios.

Hormonal profile changes induced by pesticide mixture exposure in female rats revealed by hair analysis.

Pesticide exposure has been associated with hormonal disruption in both animals and humans. However, there is limited knowledge of the combined effects of complex mixtures of pesticides on endogenous hormone levels. Here, we used hair analysis to assess the impact of a pesticide mixture comprising 19 components from multiple chemical classes at eight doses of 0-400μg/kg body weight (bw) three times per week per component on concentrations of 36 steroid and thyroid hormones in female rats over a 90-day exposure period. We detected 13 hormones in rat hair, namely estradiol (E2), androstenedione (AD), testosterone (T), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), pregnenolone (P5), progesterone (P4), 11-deoxycorticosterone (11-DOC), corticosterone (CORT), 3,3'-l-diiodothyronine (T2), 3,5,3'-l-triiodothyronine (T3), 3,3',5'-l-triiodothyronine (rT3), and 3,5,3',5'-l-tetraiodothyronine (T4). In comparison to the control group, hair E2 concentration was significantly lower in the two highest (200 and 400μg/kg bw) exposure groups, whereas hair DHEAS and CORT concentrations were significantly higher in the 40μg/kg bw and the highest exposure groups, respectively. Results from generalized additive models suggest that pesticide exposure resulted in monotonic dose responses in hair E2 concentration, CORT concentration and DHEA/CORT molar ratio but nonmonotonic dose responses in hair T concentration, DHEAS concentration, P4/P5 and DHEA/DHEAS molar ratios. The associations of E2, CORT and DHEA/CORT ratio with exposure intensity were confirmed by their significant linear relationships with hair concentrations of at least 23 of the 25 exposure biomarkers analyzed. Our results demonstrate that exposure to low levels of the pesticide mixture evaluated here can alter hair reproductive and adrenal hormones levels.

Effects of long-term direct exposure to pesticides towards immune function: a cross-sectional study

Exposure to pesticides is an important risk factor for genotoxicity in farmworkers. Once there is a possibility that genotoxicity is related to the autoimmunity process, the present study aimed to characterize genotoxicity and possible immunological alterations related to chronic and direct exposure to a complex mixture of pesticides. It employed Interleukin–6 (IL-6), Interferon-gamma (IFN-γ), IL-12 and IL-10 as biomarkers of cellular immune function. DNA damage was greater in the exposed group (p < 0.05). Both IL-6 and IL-12 were significantly increased in the exposed group (p < 0.05 and < 0.01, respectively); on the other hand, Il-10 levels were decreased (p < 0.01). Evaluation of the association between cytokine levels and epidemiological variables showed a correlation between alcohol consumption and higher levels of IL-12 (p < 0.05). In addition, negative correlation between % DNA and IFN-γ (p < 0.05) as well as % DNA and IL-6 (p < 0.05) and positive correlation was found between IL-6 levels and absolute number of T CD8+ lymphocytes (p < 0.05) as well as between IL-10 levels and absolute number of T CD4+ lymphocytes (p < 0.05). No association between all the evaluation parameters was found in the non-exposed group. The observed alterations related to chronic and direct exposure to a complex mixture of pesticides may allow the monitoring of these workers to minimize the negative impact caused on their health.

Mix-Tool: An Edge-of-Field Approach to Predict Pesticide Mixtures of Concern in Surface Water From Agricultural Crops.

Current regulation on the authorization of plant protection products (PPPs) in the European Union is limited to the evaluation of ecological risks for the single active substances they contain. However, plant protection treatments in agriculture often consist of PPPs already containing more than one active substance; moreover, each cropped field receives multiple applications per year, leading to complex pesticide mixtures in the environment. Different transport processes lead to a multitude of heterogeneous and potentially toxic substances that, for example, may reach water bodies and act simultaneously on natural freshwater ecosystems. In this context, the development of methodologies and tools to manage risks of pesticides mixtures is imperative to improve the current ecological risk assessment procedures and to avoid further deterioration of ecological quality of natural resources. The present study suggests new procedures for identifying pesticide mixtures of potential concern released from agricultural crops in surface water. The approach follows the European Union regulatory context for the authorization of PPPs in the market (edge‐of field risk assessment) and requires the use of Forum for the Co‐ordination of pesticide fate models and their Use (FOCUS) models (Step 3 and 4) for calculating the concentrations in surface water of mixture components on a daily basis. Moreover, it uses concentration addition models to calculate the toxic potency of the pesticide mixtures released by a treated crop. To implement this procedure, we developed a simple Microsoft‐Excel‐based tool. We also considered two case studies (maize and apple tree), representative of Italian agricultural scenarios for annual and perennial crops. Moreover, we compared results with 3 years of monitoring data of surface water bodies of the Lombardia region (northern Italy) where the two crops are largely present. Environ Toxicol Chem 2022;41:2028–2038. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Survival analysis of African catfish and Nile tilapia briefly exposed to complex pesticide mixtures

BackgroundPulse exposures are the consequences of the intermittent release of pollutants in the environment. Brief exposure of aquatic organisms to high concentrations of pesticides simultaneously occurs, particularly in small watercourses during high flows. The effects of pulse exposure often include effects occurring during and after the exposure. Despite this, routine toxicity tests procedures often ignore brief exposure scenarios and the role of time in toxicity. We conducted a pulse toxicity test by briefly exposing African catfish and Nile tilapia fingerlings to pesticide mixtures of atrazine, mancozeb, chlorpyrifos, and lambda-cyhalothrin. The study aimed to estimate pesticide mixture interaction in pulse-exposed fish and elucidate the influence of species differences on the response of fish to the pesticide mixture.ResultsDespite the similarity in fingerlings weight, African catfish had a significantly higher survival probability than Nile tilapia after exposure to atrazine-mancozeb mixture. However, the survival probability of African catfish and Nile tilapia fingerlings were similar after exposure to atrazine-chlorpyrifos, atrazine-lambda cyhalothrin, mancozeb-chlorpyrifos, mancozeb-lambda cyhalothrin, chlorpyrifos-lambda cyhalothrin, and quaternary mixture (p > 0.05). The survival probability of exposed fingerlings was significantly lower for continuous than pulse exposure to the mixtures (p < 0.01). Nevertheless, the survival probability of 60 min of pulse exposure to 13.49 mg/L mancozeb-lambda cyhalothrin was similar to continuous exposure for 96 h. Atrazine-mancozeb, atrazine-chlorpyrifos, atrazine-lambda cyhalothrin, mancozeb-chlorpyrifos, mancozeb-lambda cyhalothrin, and the quaternary pesticide mixture were antagonists in African catfish but not in Nile tilapia. At the same time, chlorpyrifos-lambda-cyhalothrin was antagonistic in Nile tilapia but not African catfish.ConclusionsPesticide mixture interaction was antagonist but specie-dependent. Innate intrinsic and extrinsic deterministic factors and, to a limited extent, stochastic processes may have influenced the survival probability of African catfish, and Nile tilapia pulsed exposed to complex pesticide mixtures. Pulse toxicity assessment using survival analysis is relevant in ecotoxicology as it enables the study of factors that can influence pulse toxicity.

Organic Matter Processing on Dry Riverbeds is More Reactive to Water Diversion and Pollution Than on Wet Channels

Rivers are severely affected by human activities and many are simultaneously impacted by multiple stressors. Water diversion for hydropower generation affects ecosystem functioning of the bypassed reaches, which can alternate between periods with natural discharge and others with reduced flow that increase the surface of dry riverbeds. In parallel, urban pollution contributes a complex mixture of nutrients, organic matter, heavy metals, pesticides, and drugs, thus becoming an important stressor in rivers. However, there is little information on the interaction between both stressors on ecosystem functioning and, particularly, on organic matter processing, a key process linked to the input of energy to food webs. To assess the impact of water diversion and urban pollution on organic matter processing, we selected four rivers in a pollution gradient with a similar diversion scheme and compared reaches upstream and downstream from the diversion weirs. We measured leaf-litter decomposition and carbon dioxide (CO2) fluxes in both the wet channel and the dry riverbed. Water diversion and pollution in the wet channel did not affect CO2 fluxes but reduced microbial decomposition, whereas in the dry riverbed, their interaction reduced total and microbial decomposition and CO2 fluxes. Thus, both stressors affected organic matter processing stronger in dry riverbeds than in the wet channel. These results show that dry riverbeds must be taken into account to assess and manage the impacts of human activities on river ecosystems.

Estimation of pesticide mixture interaction in Nile tilapia (Oreochromis niloticus) using survival analysis

The acute toxicity of the pesticides atrazine, mancozeb, chlorpyrifos and lambda-cyhalothrin, acting singly and jointly, was assessed on Nile tilapia (Oreochromis niloticus) fingerlings. Median lethal concentration (LC50), median lethal time (LT50), and mixture interaction were estimated, whereas survival analysis was used to model time-to-death. The most toxic single and joint mixture was lambda-cyhalothrin and chlorpyrifos-lambda cyhalothrin, respectively. The risk of death (RoD) of fingerlings exposed to 9.22 mg l−1 atrazine-mancozeb mixture was 1.76 times higher than fingerlings exposed to 9.0 mg l−1 atrazine (p > 0.05). However, RoD of fingerlings exposed to 9.95 mg l−1 atrazine-chlorpyrifos was 5.59 times higher than fingerlings exposed to 9.0 mg l−1 atrazine (p < 0.01). The risk of death of fingerlings exposed to 20.8 mg l−1 atrazine-lambda cyhalothrin was 2.81 times higher than 21.0 mg l−1 atrazine. The toxicity of 2.3 mg l−1 mancozeb-chlorpyrifos was 254.25 higher than 2.2 mg l−1 mancozeb (p < 0.01). Fingerlings exposed to 4.33 mg l−1 mancozeb-lambda cyhalothrin and 0.177 mg l−1 chlorpyrifos-lambda cyhalothrin mixture were 0.02 and 0.14 times less likely to die than those exposed to 4.8 mg l−1 mancozeb and 0.17 mg l−1 chlorpyrifos, respectively (p < 0.01). Atrazine-mancozeb, atrazine-chlorpyrifos, atrazine-lambda cyhalothrin, and mancozeb-chlorpyrifos interaction were synergistic, and their relative risk was >1. Both mancozeb-lambda cyhalothrin and lambda-cyhalothrin-chlorpyrifos mixtures were antagonistic, and their relative risk was less than 1. Survival analysis can show interaction in complex pesticide mixtures.

Agricultural surface water, imidacloprid, and chlorantraniliprole result in altered gene expression and receptor activation in Pimephales promelas

The toxicity of single pesticides is likely underestimated when considering complex pesticide mixtures found in agricultural runoff and this is especially true for newer pesticides with little toxicity data on non-target species. The goal of our study was to compare the toxicity of two newer pesticides, imidacloprid (IMI) and chlorantraniliprole (CHL), when an invertebrate and fish were exposed to single compounds, binary mixtures or surface water collected near agricultural fields. A secondary goal was to determine whether changes in select subcellular molecular pathways correspond to the insecticides’ mechanisms of activity in aquatic organisms. We conducted acute (96 h) exposures using a dilution series of field water and environmentally relevant concentrations of single and binary mixtures of IMI and CHL. We then evaluated survival, gene expression and the activity of IMI toward the n-acetylcholine receptor (nAChR) and CHL activity toward the ryanodine receptor (RyR). Both IMI and CHL were detected at all sampling locations for May 2019 and September 2019 sampling dates and exposure to field water led to high invertebrate but not fish mortality. Fish exposed to field collected water had significant changes in the relative expression of genes involved with detoxification and neuromuscular function. Exposure of fish to single compounds or binary mixtures of IMI and CHL led to increased relative gene expression of RyR in fish. Furthermore, we found that IMI targets the nAChR in aquatic invertebrates and that CHL can cause overactivation of the RyR in invertebrates and fish. Overall, our finding suggests that IMI and CHL may impact neuromuscular health in fish. Expanding monitoring efforts to include sublethal and molecular assays would allow the detection of subcellular level effects due to complex mixtures present in surface water near agricultural areas.

Residential Proximity and Cumulative Risk Assessment of Agricultural Pesticide Applications in California

BACKGROUND AND AIM: Pesticide exposure based on residential proximity to agricultural applications has been associated with adverse health outcomes including developmental neurotoxicity such as reduced IQ and autism spectrum disorder, adverse birth outcomes such as low birth weight and birth defects as well as increased risk of cancer. However, epidemiological studies have primarily assessed individual pesticides or groups of pesticides sharing a common mechanism of action. In reality, individuals living near agricultural pesticide applications are exposed to a complex mixture of pesticides impacting a broad range of potential health harms. Here we develop a toxicity index for pesticide mixtures based on multiple chronic toxicity endpoints, and identify residential areas at greatest risk of adverse health outcomes due to pesticide exposure. METHODS: Data for pesticide applications were downloaded from the California Pesticide Use Reporting program (PUR). Over 400 active ingredients were assessed for chronic toxicity endpoints including carcinogenicity, neurotoxicity, endocrine disruption, thyroid toxicity and respiratory toxicity based on classifications from authoritative bodies including the EPA, IARC, and EFSA. We developed a toxicity index for each active ingredient based on the severity of the health outcome, the strength of the association, the reliability of the data source and the NOAEL if available. We then combined the toxicity index with application rates for all pesticides applied within a square mile. RESULTS:Pesticides with the greatest toxicity index include active ingredients previously associated with adverse health outcomes including organophosphates and carbamates. Other pesticides with high toxicity indices include neonicotinoids, pyrethroids and triazole fungicides. Residential areas of high risk of potential adverse health effects due to pesticide exposure from nearby agricultural applications were identified. CONCLUSIONS:Using the pesticide toxicity index and application rates in combination with residential proximity agricultural applications can help identify and prioritize high risk areas for future pesticide policy actions to best protect public health. KEYWORDS: pesticides, cumulative risk assessment, residential proximity

Risk from pesticide mixtures – The gap between risk assessment and reality

Pesticide applications in agricultural crops often comprise a mixture of plant protection products (PPP), and single fields face multiple applications per year leading to complex pesticide mixtures in the environment. Restricted to single PPP, the current European Union PPP regulation, however, disregards the ecological risks of pesticide mixtures. To quantify this additional risk, we evaluated the contribution of single pesticide active ingredients to the additive mixture risk for aquatic risk indicators (invertebrates and algae) in 464 different PPP used, 3446 applications sprayed and 830 water samples collected in Central Europe, Germany. We identified an average number of 1.3 different pesticides in a single PPP, 3.1 for complete applications often involving multiple PPP and 30 in stream water samples. Under realistic worst-case conditions, the estimated stream water pesticide risk based on additive effects was 3.2 times higher than predicted from single PPP. We found that in streams, however, the majority of regulatory threshold exceedances was caused by single pesticides alone (69% for algae, 81% for invertebrates). Both in PPP applications and in stream samples, pesticide exposure occurred in repeated pulses each driven by one to few alternating pesticides. The time intervals between pulses were shorter than the 8 weeks considered for ecological recovery in environmental risk assessment in 88% of spray series and 53% of streams. We conclude that pesticide risk assessment should consider an additional assessment factor to account for the additive, but also potential synergistic simultaneous pesticide mixture risk. Additionally, future research and risk assessment need to address the risk from the frequent sequential pesticide exposure observed in this study.

Assessment of Cytogenetic Damage and Cholinesterases' Activity in Workers Occupationally Exposed to Pesticides in Zamora-Jacona, Michoacan, Mexico.

Pesticides have been considered as potential chemical mutagens; however, little is known about toxic and genotoxic effects during pesticide application in Zamora-Jacona, Michoacan State in Mexico. This study sought to determine DNA damage and cholinesterase activities inhibitions in 54 agricultural workers exposed to complex mixtures of pesticides vs. control group (26 individuals) using Comet assay in peripheral whole blood, micronucleus (MN) test in oral mucosa cells, Cytokinesis-blocked MN assay in lymphocytes (L-CBMNcyt) and measuring AChE and BChE activities in whole blood and plasma samples, respectively. Exposed subjects demonstrated significantly elevated levels of primary (Comet assay: tail intensity, tail length, tail moment, Olive tail moment) and permanent DNA damage (MN assay: in blood/buccal cells; frequencies of nuclear buds, binucleated cells, cells with condensed chromatin, karyorrhexis, pyknosis, and karyolysis). However, inhibition of cholinesterase activities (AChE and BChE) was not observed in the workers. Confounding factors including sex, age, BMI, working exposure period, protection level, smoking habit (cigarettes per day units), alcohol consumption (weekly), medication, were considered in the analysis. These combined techniques demonstrated usefulness in the health hazards risks pesticide exposure assessment and suggested the need for periodic monitoring together with the education and the training of occupational workers for the safe application of potentially harmful pesticides.

&lt;i&gt;In sit&lt;/i&gt; assessment of genotoxicity in tadpoles (Amphibia: Anura) in impacted and protected areas of Chapada Diamantina, Brazil

We evaluated the genotoxic potential of surface water bodies in an area impacted by agrochemicals, in the Ibicoara-Mucugê agropole, Bahia and those within protected areas in Chapada Diamantina, Bahia, Brazil, using the micronucleus test as a biomarker in tadpole species native neotropicals. Our hypothesis is that the closer to agricultural farms, the more affected by mutagenic and genotoxic substances will be the environment and resident organisms. The micronucleus frequencies were determined in tadpoles of 11 species collected in 11 surface water bodies in coffee, corn, vegetable, and pumpkin plantations, as well as in adjacent sites within a protection area. Tadpoles of seven species found in agricultural sites exhibited higher frequencies of micronuclei compared to the six species from the protected locations. The species Bokermannohyla oxente and Leptodactylus latrans collected in agricultural areas exhibited the highest frequencies of micronucleated erythrocytes compared to the populations from the protected areas (p &lt;0.05). Our results indicate that compounds present in surface water bodies in agricultural areas of the Ibicoara-Mucugê farm have genotoxic effects on anurans, showing the stress responses caused by agricultural activities associated with the use of complex mixtures of pesticides. Thus, anurans are excellent sentinel species in environmental monitoring, previously indicating the effect of xenobiotics on the environment.

Risk Assessment of Pesticide Mixtures in Australian Rivers Discharging to the Great Barrier Reef.

Rivers discharging to the Great Barrier Reef carry complex pesticide mixtures. Here we present a first comprehensive ecotoxicological risk assessment using species sensitivity distributions (SSDs), explore how risk changes with time and land use, and identify the drivers of mixture risks. The analyzed data set comprises 50 different pesticides and pesticide metabolites that were analyzed in 3741 samples from 18 river and creek catchments between 2011 and 2016. Pesticide mixtures were present in 82% of the samples, with a maximum of 23 pesticides and a median of five compounds per sample. Chemical-analytical techniques were insufficiently sensitive for at least seven pesticides (metsulfuron-methyl, terbutryn, imidacloprid, clothianidin, ametryn, prometryn, and thiamethoxam). The classical mixture concepts of concentration addition and independent action were applied to the pesticide SSDs, focusing on environmental threshold values protective for 95% of the species. Both concepts produced almost identical risk estimates. Mixture risk was therefore finally assessed using concentration addition, as the sum of the individual risk quotients. The sum of risk quotients ranges between 0.05 and 122 with a median of 0.66. An ecotoxicological risk (i.e., a sum of individual risk quotients exceeding 1) was indicated in 38.5% of the samples. Sixteen compounds accounted for 99% of the risk, with diuron, imidacloprid, atrazine, metolachlor, and hexazinone being the most important risk drivers. Analysis of land-use patterns in catchment areas showed an association between sugar cane farming and elevated risk levels, driven by the presence of diuron.

Removal of triazines, triazoles and organophophates in biomixtures and application of a biopurification system for the treatment of laboratory wastewaters

Biopurification systems (BPS) have been barely explored for removing complex mixtures of pesticides. In this study, the potential of a biomixture to remove simultaneously a mixture of herbicides (triazines), fungicides (triazoles) and insecticides (organophosphates) is presented. Also, a BPS using the same biomixture was used for treating a pesticide testing laboratory wastewater containing a mixture of 38 compounds. Ecotoxicological assays were conducted on the BPS elutriates to investigate the mixture detoxification. A mixture (concentrations of 4–8 mg kg−1) run in small–scale biomixture systems (SSB) for 128 d showed 59.3% removal of triazines, 68.5% of organophosphates and no elimination of triazoles. The treatment of the laboratory wastewater (individual concentrations range: 0.0036–0.25 mg kg−1) in the pilot-scale BPS for 281 d resulted in the elimination pattern of organophosphates (90.0%) > triazoles (73.4%) > carbamates (71.3%) > triazines (54.3%). Complete detoxification towards Daphnia magna and partial detoxification in Lactuca sativa seeds germination occurred in the BPS. Although the pesticide mixture complexity is higher in the BPS, the lower concentrations found in this matrix, could explain removal differences between SSB and BPS and the apparent inhibition in the elimination of carbamates and some triazines observed in the latter. These findings suggest that disposal of pesticide–containing laboratory–wastewater should be done in separate containers, according to chemical groups before their treatment in separate BPS, in order to reduce treatment periods. Monitoring the treatment process in the BPS with a battery of ecotoxicological tests is strongly recommended.