Although microplastics (MPs) are ubiquitous in agricultural soil, little is known about the effects of MPs combined with pesticides on soil organisms and their biogenic transport through the soil profile. In this study, we conducted mesocosm experiments to observe the effects of microplastics (polyethylene (LDPE-MPs) and biodegradable microplastics (Bio-MPs)) and chlorpyrifos (CPF) on earthworm (Lumbricus terrestris) mortality, growth and reproduction, as well as the biogenic transport of these contaminants through earthworm burrows. The results showed that earthworm reproduction was not affected by any treatment, but earthworm weight was reduced by 17.6% and the mortality increased by 62.5% in treatments with 28% Bio-MPs. Treatments with 28% LDPE-MPs and 7% Bio-MPs combined with CPF showed greater toxicity while the treatment with 28% Bio-MPs combined with CPF showed less toxicity on earthworm growth as compared to treatments with only MPs. The treatments with 1250 g ha−1 CPF and 28% Bio-MPs significantly decreased the bioaccumulation of CPF in earthworm bodies (1.1 ± 0.2%, w w−1), compared to the treatment with CPF alone (1.7 ± 0.4%). With CPF addition, more LDPE-MPs (8%) were transported into earthworm burrows and the distribution rate of LDPE-MPs in deeper soil was increased. No effect was observed on the transport of Bio-MPs. More CPF was transported into soil in the treatments with LDPE-MPs and Bio-MPs, 5% and 10% of added CPF, respectively. In addition, a lower level of the CPF metabolite 3,5,6-trichloropyridinol was detected in soil samples from the treatments with MPs additions than without MP additions, indicating that the presence of MPs inhibited CPF degradation. In conclusion, Bio-MPs caused significant toxicity effects on earthworms and the different types of MPs combined with CPF affected earthworms differently, and their transport along the soil profile. Thus, further research is urgently needed to understand the environmental risks of MPs and MP-associated compounds in the soil ecosystem.

Chlorpyrifos (CPF), as an organophosphate insecticide extensively used in the modern agricultural system, has been gradually banned in many countries due to its reported health risks to organisms, including humans. This study used simulated paddy field experiments and carbon-14 tracing to explore the possibility of reducing environmental risks of chlorpyrifos application through appropriate agronomic practice. Results showed ¹⁴C-CPF concentration in rice plants planted in the red soil (RS) was significantly higher than that in black soil (BS) and fluvo-aquic soil (FS). The application of biochar and chicken manure in RS reduced ¹⁴C-CPF accumulation in rice plants, and the content of ¹⁴C-CPF in rice grains decreased by 25% and 50%, respectively. Adding biochar to all three soils reduced the migration of ¹⁴C-CPF, especially in FS with the highest risk of ¹⁴C-CPF migration. The addition of chicken manure in FS reduced the migration of ¹⁴C-CPF and the total residual amount of ¹⁴C-CPF in the soil. In addition, chicken manure treatment increased the formation of ¹⁴C-bound residues (BRs) in soils and changed the distribution ¹⁴C-BRs in humus. The results indicated that the degree of environmental risks associated with the CPF application varies with soil types and could be reduced by introducing suitable exogenous organic matter into different soils, which is of great significance for guiding the scientific application of chlorpyrifos in agronomic practices.

A monitoring network was established in streams within a catchment near the Costa Rican Pacific coast (2008–2011) to estimate the impact of pesticides in surface water (84 samples) and sediments (84 samples) in areas under the influence of melon and watermelon production. A total of 66 (water) and 47 (sediment) pesticides were analyzed, and an environmental risk assessment (ERA) was performed for four taxa (algae, Daphnia magna, fish and Chironomus riparius). One fungicide and seven insecticides were detected in water and/or sediment; the fungicide azoxystrobin (water) and the insecticide cypermethrin (sediments) were the most frequently detected pesticides. The insecticides endosulfan (5.76 μg/L) and cypermethrin (301 μg/kg) presented the highest concentrations in water and sediment, respectively. The ERA revealed acute risk in half of the sampling points of the melon-influenced area and in every sampling point from the watermelon-influenced area. Safety levels were exceeded within and around the crop fields, suggesting that agrochemical contamination was distributed along the catchment, with potential influence of nearby crops. Acute risk was caused by the insecticides chlorpyrifos, cypermethrin and endosulfan to D. magna, fish and C. riparius; the latter was the organism with the overall highest/continuous risk. High chronic risk was determined in all but one sampling point, and revealed a higher number of pesticides of concern. Cypermethrin was the only pesticide to pose chronic risk for all benchmark organisms. The results provide new information on the risk that tropical crops pose to aquatic ecosystems, and highlight the importance of including the analysis of sediment concentrations and chronic exposure in ERA.

Novel stressors introduced by human activities increasingly threaten freshwater ecosystems. The annual application of more than 2.3 billion kg of pesticide active ingredient and 22 billion kg of road salt has led to the contamination of temperate waterways. While pesticides and road salt are known to cause direct and indirect effects in aquatic communities, their possible interactive effects remain widely unknown. Using outdoor mesocosms, we created wetland communities consisting of zooplankton, phytoplankton, periphyton, and leopard frog (Rana pipiens) tadpoles. We evaluated the toxic effects of six broad-spectrum insecticides from three families (neonicotinoids: thiamethoxam, imidacloprid; organophosphates: chlorpyrifos, malathion; pyrethroids: cypermethrin, permethrin), as well as the potentially interactive effects of four of these insecticides with three concentrations of road salt (NaCl; 44, 160, 1600 Cl⁻ mg/L). Organophosphate exposure decreased zooplankton abundance, elevated phytoplankton biomass, and reduced tadpole mass whereas exposure to neonicotinoids and pyrethroids decreased zooplankton abundance but had no significant effect on phytoplankton abundance or tadpole mass. While organophosphates decreased zooplankton abundance at all salt concentrations, effects on phytoplankton abundance and tadpole mass were dependent upon salt concentration. In contrast, while pyrethroids had no effects in the absence of salt, they decreased zooplankton and phytoplankton density under increased salt concentrations. Our results highlight the importance of multiple-stressor research under natural conditions. As human activities continue to imperil freshwater systems, it is vital to move beyond single-stressor experiments that exclude potentially interactive effects of chemical contaminants.

Given extensive use of pesticides in agriculture, there is concern for unintended consequences to non-target species. The non-target freshwater amphipod, Hyalella azteca has been found to show resistance to the organophosphate (OP) pesticide, chlorpyrifos, resulting from an amino acid substitution in acetylcholinesterase (AChE), suggesting a selective pressure of unintended pesticide exposure. Since resistant organisms can survive in contaminated habitats, there is potential for them to accumulate higher concentrations of insecticides, increasing the risk for trophic transfer. In the present study, we estimated the uptake and elimination of chlorpyrifos in non-resistant US Lab, and resistant Ulatis Creek (ULC Resistant), H. azteca populations by conducting 24-h uptake and 48-h elimination toxicokinetic experiments with ¹⁴C-chlorpyrifos. Our results indicated that non-resistant H. azteca had a larger uptake clearance coefficient (1467 mL g⁻¹ h⁻¹) than resistant animals (557 mL g⁻¹ h⁻¹). The half-life derived from the toxicokinetic models also estimated that steady state conditions were reached at 13.5 and 32.5 h for US Lab and ULC, respectively. Bioaccumulation was compared between non-resistant and resistant H. azteca by exposing animals to six different environmentally relevant concentrations for 28 h. Detection of chlorpyrifos in animal tissues indicated that resistant animals exposed to high concentrations of chlorpyrifos were capable of accumulating the insecticide up to 10-fold higher compared to non-resistant animals. Metabolite analysis from the 28-h concentration experiments showed that between 20 and 50 % parent compound was detected in H. azteca. These results imply that bioaccumulation potential can be more significant in chlorpyrifos resistant H. azteca and may be an essential factor in assessing the full impacts of toxicants on critical food webs, especially in the face of increasing pesticide and chemical runoff.

Large-scale ecotoxicological studies have technical and ethical limitations, both related to the need to expose large numbers of individuals to potentially harmful compounds. The computational modeling is a complementary useful and predictive tool that overcomes these limitations. Considering the increasing interest in the effects of pesticides on behavioral traits, the aim of this study was to evaluate the effects of chlorpyrifos (CPF) on intra- and inter-specific interactions of anuran tadpoles, complementing traditional ecotoxicological tools with a theoretical analysis verified by computational simulations. Experiments were developed under two consecutive phases: a first phase of exposure (treated and control group), and a second phase of interactions. The second phase consisted of evaluating the effects of CPF on intra- and inter-specific interactions of exposed C. ornata (Co) tadpoles acting as predators and unexposed Rhinella fernandezae (Rf) tadpoles acting as prey (Experiment I), under different predator-prey proportions (0/10 = 0Co–10Rf, 2/8, 4/6, 6/4, 8/2, 10/0). Also, intraspecific interactions of three Co tadpoles under different conditions of exposure were evaluated (Experiment II: 3 exposed Co, 2 exposed Co/1 non-exposed, 1 exposed Co/2 non-exposed). During the exposure phase, chlorpyrifos induced significant mortality from 48 h (48 h: p < 0.05, 72 h–96 h: p < 0.001), irregular swimming, tail flexure, and the presence of subcutaneous air. Also, it induced effects on the sounds emitted after 96 h of exposure, registering a smaller number of pulses and higher dominant frequencies, and altered intra- and inter-specific interactions. During the interaction phase, the larvae continued to show sound effects, however, the antipredator mechanism continued to be operating and efficient. Finally, it was possible to model the behavior of the larvae under the effects of chlorpyrifos. We conclude that experimental data and computational modeling matched. Therefore, computational simulation is a valuable ecotoxicological tool that provides new information and allows prediction of natural processes.

While extreme high temperatures are an important aspect of global warming, their effects on organisms are relatively understudied, especially in ecotoxicology. Sequential exposure to heat spikes and pesticides is a realistic scenario as both are typically transient stressors and are expected to further increase in frequency under global warming. We tested the effects of exposure to a lethal heat spike and subsequently to an ecologically relevant lethal pulse exposure of the pesticide chlorpyrifos in the larvae of mosquito Culex pipiens. The heat spike caused direct and delayed mortality, and resulted in a higher heat tolerance and activity of acetylcholinesterase, and a lower fat content in the survivors. The chlorpyrifos exposure caused mortality, accelerated growth rate, and decreased the heat tolerance and the activity of acetylcholinesterase. The preceding heat spike did not change how chlorpyrifos reduced the heat tolerance. Notably, the preceding heat spike did lower the lethal effect of the pesticide, which makes an important novel finding at the interface of ecotoxicology and global change biology, and adds a new dimension to the “climate-induced toxicant sensitivity” (CITS) concept. This may be due to both survival selection and cross-tolerance, and therefore likely a widespread phenomenon. Our results emphasize the importance of including extreme high temperatures as an important transient global change stressor in ecotoxicology.

Water pollution by pesticides used in agriculture is currently a major concern both in Spain and in Europe as a whole, prompting the need to evaluate water quality and ecological risk in areas of intensive agriculture. This study involved monitoring pesticide residues and certain degradation products in surface and ground waters of the Denomination of Origin (DO) Jumilla vineyard area in Spain. Sixty-nine pesticides were selected and evaluated at twenty-one sampling points using a multi-residue analytical method, based on solid-phase extraction (SPE) and analysis by liquid chromatography coupled with mass spectrometry (LC-MS), providing reliable results. Twenty-six compounds from those selected were detected in the samples analyzed (eleven insecticides including one degradation product, nine herbicides, and six fungicides) and fifteen of them were found in concentrations over 0.1 μg L⁻¹ (upper threshold established by the EU for pesticides detected in waters for human consumption). Indoxacarb was present in more than 70% of the samples, being the most frequently detected compound in water samples. Some pesticides were ubiquitous in all the water samples. Ecotoxicological risk indicators, toxic units (TUs) and risk quotients (RQs), for algae, Daphnia magna and fish were calculated to estimate the environmental risk of the presence of pesticides in waters. The compounds with the highest risk were the herbicides pendimethalin, with RQ values > 1 for the three aquatic organisms, and diflufenican, posing a high risk for algae and fish, and the insecticide chlorpyrifos, with a high risk for Daphnia magna and fish. The ∑TUi determined for water at each sampling point posed only a high risk for the three aquatic organisms in a sample. These results are important for considering the selection of pesticides with less environmental risk in intensive agricultural areas.

Chlorpyrifos is one of the most heavily used pesticides in domestic and agricultural insect prevention globally. Given the potential neurotoxicity of chlorpyrifos and its high detection rates in food and drinking water, health risks attributable to chlorpyrifos residue in Chinese drinking water and food in both China and Denmark were assessed in this study. Mixed left-censored handling models were used to deal with the non-detected values in chlorpyrifos concentrations. Results show that chronic exposure imputed to chlorpyrifos residue is much lower than the reference dose, and will thus not pose appreciable health risk to the consumer. Compared to the total exposure from chlorpyrifos in drinking water and food sources, chronic exposure from drinking water sources in China accounts for 0–4.4%. Health risk owing to chlorpyrifos in food within China is 6-7-fold higher than in Denmark, and this coincides with the fact that all application of chlorpyrifos is banned in Denmark, in contrast to China. However, the Danish consumers are still exposed from imported food items. The main health risk contributors in China are the food groups of Grains and grain-based products and Vegetable and vegetable products, while the main chronic health risk contributor in Denmark is the food group of imported fruit and fruit products.

Pesticide exposure is regarded as a contributing factor to the high gross loss rates of managed colonies of Apis mellifera. Pesticides enter the hive through contaminated nectar and pollen carried by returning forager honey bees or placed in the hive by beekeepers when managing hive pests. We used an in vitro rearing method to characterize the effects of seven pesticides on developing brood subjected dietary exposure at worse-case environmental concentrations detected in wax and pollen. The pesticides tested included acaricides (amitraz, coumaphos, fluvalinate), insecticides (chlorpyrifos, imidacloprid), one fungicide (chlorothalonil), and one herbicide (glyphosate). The larvae were exposed chronically for six days of mimicking exposure during the entire larval feeding period, which is the worst possible scenario of larval exposure. Survival, duration of immature development, the weight of newly emerged adult, morphologies of the antenna and the hypopharyngeal gland, and gene expression were recorded. Survival of bees exposed to amitraz, coumaphos, fluvalinate, chlorpyrifos, and chlorothalonil was the most sensitive endpoint despite observed changes in many developmental and physiological parameters across the seven pesticides. Our findings suggest that pesticide exposure during larvae development may affect the survival and health of immature honey bees, thus contributing to overall colony stress or loss. Additionally, pesticide exposure altered gene expression of detoxification enzymes. However, the tested exposure scenario is unlikely to be representative of real-world conditions but emphasizes the importance of proper hive management to minimize pesticide contamination of the hive environment or simulates a future scenario of increased contamination.