Ecotoxicity and Risk to Zooplankton of a Novel Insecticide Used in Rice Paddies in a Simulated Ecosystem.

Diamide insecticides are widely used in rice paddies and pose a potential threat to aquatic organisms. However, the risk research related to their application in major rice-producing areas is very limited, especially mesocosm research to simulate the impact on aquatic ecosystems of long-term exposure, as well as exposure analysis based on local models and local scenarios. To assess potential risks from a novel diamide insecticide (tetrachlorantraniliprole) to aquatic nontarget organisms in the field over long-term exposure, an outdoor mesocosm study was performed, and the environmental concentrations were predicted by the multimedia paddy-pond model (TOPRICE). The mesocosm experiment showed that tetrachlorantraniliprole mainly stayed in the aqueous phase after entering the water body. Although the chemical dissipated quickly in the aqueous phase (half-life of 0.79-1.5 days), it showed toxic effects on zooplankton communities. Cladocerans, represented by Simocephalus vetulus, were most sensitive to tetrachlorantraniliprole stress. Significant short-term toxicity to cladocerans occurred in all treatment groups, but all recovered within 8 weeks except for the highest concentration group (30.0 µg /L). Based on the ecological recovery results, 7.74 µg tetrachlorantraniliprole/L (nominal concentration, 10.0 µg /L) is suggested to be the no-observed-ecological-adverse-effect concentration (NOEAEC) for the zooplankton community. When this NOEAEC was compared with predicted environmental concentrations (PECs; the PECs in natural ponds simulated by the TOPRICE model for 148 application scheme combinations in major rice-producing areas), a relatively high risk of applying tetrachlorantraniliprole during the rice tillering stage was found. The present study makes a positive contribution to the hypothesis that the current Tier 1 approaches for global acute risk assessment have a sufficient protective effect for assessing the risk of tetrachlorantraniliprole to aquatic organisms. Also, the present results should help us to gain a fuller understanding of the ecological risk of diamide insecticides in aquatic ecosystems and their rational application schemes. Environ Toxicol Chem 2024;43:429-439. © 2023 SETAC.

Nitrogen-Doped Hierarchical Porous Carbon Derived from Coal for High-Performance Supercapacitor.

The surface properties and the hierarchical pore structure of carbon materials are important for their actual application in supercapacitors. It is important to pursue an integrated approach that is both easy and cost-effective but also challenging. Herein, coal-based hierarchical porous carbon with nitrogen doping was prepared by a simple dual template strategy using coal as the carbon precursor. The hierarchical pores were controlled by incorporating different target templates. Thanks to high conductivity, large electrochemically active surface area (483 m2 g-1), hierarchical porousness with appropriate micro-/mesoporous channels, and high surface nitrogen content (5.34%), the resulting porous carbon exhibits a high specific capacitance in a three-electrode system using KOH electrolytes, reaching 302 F g-1 at 1 A g-1 and 230 F g-1 at 50 A g-1 with a retention rate of 76%. At 250 W kg-1, the symmetrical supercapacitor assembled at 6 M KOH shows a high energy density of 8.3 Wh kg-1, and the stability of the cycling is smooth. The energy density of the symmetric supercapacitor assembled under ionic liquids was further increased to 48.3 Wh kg-1 with a power output of 750 W kg-1 when the operating voltage was increased to 3 V. This work expands the application of coal-based carbon materials in capacitive energy storage.

Dual-Salt-Induced Hierarchical Porous Structure in a Carbon Sheet for High Performance Supercapacitors.

Porous carbon, one of the characteristic materials for electrochemical energy storage devices, has been paid wide-ranging attention. However, balancing the reconcilable mesopore volume with a large specific surface area (SSA) was still a challenge. Herein, a dual-salt-induced activation strategy was developed to obtain a porous carbon sheet with ultrahigh SSA (3082 m2 g-1), desirable mesopore volume (0.66 cm3 g-1), nanosheet morphology, and high surface O (7.87%) and S (4.0%) content. Hence, as a supercapacitor electrode, the optimal sample possessed a high specific capacitance (351 F g-1 at 1 A g-1) and excellent rate performance (holding capacitance up to 72.2% at 50 A g-1). Furthermore, the assembled zinc-ion hybrid supercapacitor also exhibited superior reversible capacity (142.7 mAh g-1 at 0.2 A g-1) and highly stable cycling (71.2 mAh g-1 at 5 A g-1 after 10,000 cycles with retention of 98.9%). This work was delivered a new possibility for the development of coal resources for the preparation of high performance porous carbon materials.

Bioaccumulation and toxicity of terbuthylazine in earthworms (Eisenia fetida).

Terbuthylazine is an effective and widely used s-triazine herbicide. However, limited data exists on its toxicity and bioaccumulation in earthworms (Eisenia fetida). In this study, we investigated the bioaccumulation, antioxidant enzyme activity, detoxification enzyme activity, and DNA damage in earthworms when exposed to terbuthylazine. The results indicated that terbuthylazine in soil had low bioaccumulation in earthworms and the biota-soil accumulation factors of terbuthylazine declined with an increasing soil terbuthylazine concentration. In the enzyme activity assays, the superoxide dismutase (SOD), catalase (CAT), and glutathione-S-transferase (GST) activities showed upward trends when compared with the control. The carboxylesterase (CarE) activity increased on day 21. The 8-hydroxy-2-deoxyguanosine (8-OHdG) content, a DNA damage bioindicator, was higher than that of the control on day 21. Combined with the integrated biological response index version 2 analysis, these results can provide a comprehensive evaluation of the toxicological effects that terbuthylazine has on earthworms and soil ecosystems.

Evaluation of the predictive values of elevated serum L-homoarginine and dimethylarginines in preeclampsia.

L-homoarginine (hARG) is involved in nitric oxide biosynthesis, but its role and concentration in preeclampsia (PE) have not been fully revealed. The purpose of this study was to develop and validate a feasible clinical assay to quantify serum hARG, arginine (ARG), asymmetric (ADMA) and symmetric dimethylarginines (SDMA) levels by LC-MS/MS and investigate their differences at different stages of pregnancy with or without preeclampsia. Serum samples were collected from 84 pregnant women without complications (controls), 84 with mild preeclampsia (MPE), and 81 with severe preeclampsia (SPE) at various gestation stages (before the 20th week, during the 20th-28th week or after the 28th week of gestation). No significant difference in ARG levels was observed between PE and controls at any stage (P > 0.05). The serum hARG levels and hARG/ADMA ratios of MPE before the 20th week were higher than those of controls (P < 0.001). ADMA levels of MPE were higher than those of controls during the 20th-28th week (P < 0.01). SDMA levels of SPE were higher than those of MPE (P < 0.01) and controls (P < 0.05) after the 28th week. Elevated serum hARG before the 20th week was identified as an independent predictor for PE (OR = 1.478, 95% CI 1.120-1.950). ROC curve analysis showed serum hARG before the 20th week had a good potential to predict MPE (AUC = 0.875, 95% CI 0.759-0.948). In conclusion, our study indicated that elevated serum hARG and dimethylarginine levels detected by LC-MS/MS might serve as potential biomarkers for the early prediction of PE.

Joint toxic effects of cadmium and four pesticides on the earthworm (Eisenia fetida).

The contaminants, instead of single contaminants, are usually found as chemical mixtures in terrestrial environment. However, little information is now available about the joint effects of heavy metals and farm chemicals on the soil organism. Our current study aimed to assess the combined toxicity of cadmium (Cd) and four pesticides (atrazine, chlorpyrifos, lambda-cyhalothrin and abamectin) to the earthworm (Eisenia fetida) with binary, ternary, quaternary and quinquenary mixtures. Two distinct kinds of bioassay systems were applied in this study, including contact filter paper test and artificial soil test. Synergistic effects were observed from two quaternary mixtures (Cd-Atrazine-Chlorpyrifos-Lambda-cyhalothrin and Cd-Chlorpyrifos-Lambda-cyhalothrin-Abamectin) and one quinquenary mixture (Cd-Atrazine-Chlorpyrifos-Lambda-cyhalothrin-Abamectin) in both bioassay systems. Besides, two binary mixtures (Cd-Atrazine and Cd-Lambda-cyhalothrin) and three ternary mixtures (Cd-Atrazine-Chlorpyrifos, Cd-Atrazine-Lambda-cyhalothrin and Cd-Chlorpyrifos-Lambda-cyhalothrin) also exhibited synergistic effects on E. fetida by the soil toxicity test. It is more practical to evaluate the contaminant toxicities to earthworm by soil toxicity test. Therefore, the effects of chemical mixtures on soil invertebrates might be underestimated by data obtained from single toxicant. Our findings would offer a better understanding of the complex effects of chemical mixtures on non-target living creatures, and these findings provided valuable insights into the interplay of different chemicals in natural environment.

Multi-level ecotoxicological effects of imidacloprid on earthworm (Eisenia fetida).

As a neurotoxic insecticide, imidacloprid (IMI) has been widely used for crop protection. However, continuous application of such pesticide in the environment may damage the non-target organisms in soil. In the present study, we aimed to investigate the effects of IMI on earthworms in terms of survival, avoidance behavior, reproduction, detoxification enzyme activity and gene expression using a systematic experimental approach. The results showed that the 14-day LC50 value of IMI was 2.26 (2.09-2.43) mg a.i. kg-1, and the 2-day AC50 value (concentration inducing an avoidance rate of 50%) of IMI was 1.34 (1.02-1.91) mg a.i. kg-1 to E. fetida. For reproduction, the 56-day EC50 value of IMI was 0.87 (0.66-1.33) mg a.i. kg-1 to E. fetida, and there was a positive correlation between the growth rate of earthworms and the number of juveniles in IMI treatments. Activities of carboxylesterase (CarE) and glutathione-S-transferases (GST) in earthworms were disturbed by IMI exposure. Moreover, effects of IMI on the CarE activity in earthworms were more severe and sensitive compared with the GST activity. The expressions of annetocin (ann) and calreticulin (crt) at the transcriptional level were decreased upon IMI exposure, reaching the lowest levels of 0.09 fold and 0.16 fold on day 7 and day 14, respectively. Transcriptionally controlled tumor protein (tctp), heat shock protein 70 (hsp70) and gst exhibited relatively obvious variations (up-regulation or down-regulation) when the exposure duration was extended. Taken together, these results comprehensively contributed to further understandings of the impacts of IMI on earthworms.

Combined effects of four pesticides and heavy metal chromium (â ¥) on the earthworm using avoidance behavior as an endpoint.

In natural ecosystems, organisms are commonly exposed to chemical mixtures rather than individual compounds. However, environmental risk is traditionally assessed based on data of individual compounds. In the present study, we aimed to investigate the individual and combined effects of four pesticides [fenobucarb (FEN), chlorpyrifos (CPF), clothianidin (CLO), acetochlor (ACE)] and one heavy metal chromium [Cr(Ⅵ)] on the earthworm (Eisenia fetida) using avoidance behavior as an endpoint. Our results indicated that CLO had the highest toxicity to E. fetida, followed by Cr(Ⅵ), while FEN showed the least toxicity. Two mixtures of CPF+CLO and Cr(Ⅵ)+CPF+CLO+ACE exhibited synergistic effects on the earthworms. The other two quaternary mixtures of CLO+FEN+ACE+Cr(Ⅵ) and Cr(Ⅵ)+FEN+CPF+ACE at low concentrations also displayed synergistic effects on the earthworms. In contrast, the mixture of Cr(Ⅵ)+FEN had the strongest antagonistic effects on E. fetida. Besides, the quinquenary mixture of Cr(Ⅵ)+FEN+CPF+CLO+ACE also exerted antagonistic effects. These findings highlighted the importance to evaluate chemical mixtures. Moreover, our data strongly pointed out that the avoidance tests could be used to assess the effects of combined effects.

Single and joint toxicity assessment of four currently used pesticides to zebrafish (Danio rerio) using traditional and molecular endpoints.

Pesticides usually present in mixtures in surface waters, although they are traditionally regulated on an individual basis in aquatic ecosystems. In this study, we aimed to investigate the lethal and transcriptional responses of individual and combined pesticides (iprodione, pyrimethanil, pyraclostrobin and acetamiprid) on zebrafish (Danio rerio). Semi-static toxicity test indicated that the greatest toxicity to the four life stages (embryonic, larval, juvenile and adult stages) of D. rerio was detected from pyraclostrobin, followed by iprodione and pyrimethanil. In contrast, the lowest toxicity to the organisms was found from acetamiprid. Most of the selected pesticides exerted greater toxicities to D. rerio of embryonic stage compared with other life stages. Synergistic responses were observed from all binary mixtures of iprodione in combination with pyrimethanil or acetamiprid and ternary mixtures of iprodione+pyraclostrobin in combination with pyrimethanil or acetamiprid. The expressions of 16 genes related to cell apoptosis pathway, oxidative stress response, innate immunity and endocrine disruption at the mRNA level showed that zebrafish embryos were affected by the individual or combined pesticides. The expressions of P53, Tnf, TRß, Tsh and Cyp19a exhibited greater changes upon exposure to combined pesticides compared with individual pesticides. Taken together, increased toxicity might be triggered by the simultaneous presence of several pesticides in the aquatic environment, which seriously damaged the non-target organisms.

The toxic influence of paraquat on hippocampal neurogenesis in adult mice.

Paraquat, a fast-acting non-selective contact herbicide, is considered an etiological factor related to Parkinson's disease. This study investigated its effects on hippocampal neurogenesis and cognition in adult mice as well as possible mechanisms for the effects. We administered paraquat (1.25 mg/kg, intraperitoneal injection, i.p.) and an equal volume of normal saline for 3 weeks to adult male C57BL/6J mice. The results showed that hippocampus-dependent spatial learning and memory was significantly impaired in paraquat-treated mice. Moreover, paraquat administration inhibited the proliferation of neural progenitor cells, and impaired the survival and altered the fate decision of newly generated cells in the hippocampus. The expression levels of caspase-3 and glial fibrillary acidic protein were significantly higher in paraquat-treated mice than in control mice. Interestingly, paraquat reduced the phosphorylation of Akt, but did not affect the total amount of Akt. In conclusion, our findings suggest that paraquat negatively affected adult hippocampal neurogenesis and cognition function.

Elevated microRNA-520g in pre-eclampsia inhibits migration and invasion of trophoblasts.

INTRODUCTION: Pre-eclampsia (PE) is a common and severe obstetric complication. MicroRNAs (miRs) have emerged as molecules that are associated with the disease. METHODS: Quantitative reverse transcription PCR (RT-qPCR) was used for serum miR-520g characterization from 19 severe pre-eclamptic and 19 normal pregnancies. In situ hybridation was adopted to localize microRNA-520g (miR-520g). Migration and invasion of HTR-8/SVneo cells were evaluated after miR-520g mimic treatment with transwell system. MiR-520g target gene was verified in luciferase reporter system. RESULTS: The expression of serum miR-520g displayed an upward trend as pregnancies progress. At first-trimester, miR-520g in pre-eclampsia was significantly higher than that in the control, but no significant differences were found in the second and last trimesters. MiR-520g localized in cytoplasm of early trimester placental trophoblasts. The migration and invasion of HTR8/SVneo were inhibited by miR-520g mimic treatment. Matrix metalloproteinase 2 (MMP2) was verified as a direct target of miR-520g. CONCLUSIONS: Elevated maternal serum level of miR-520g level in first trimester was detected in patients with severe PE. By suppressing the migration and invasion of trophoblast via at least partial inhibition of MMP2 translation inhibition, miR-520g might play a role in the defective spiral artery remodeling, and thus contribute to pre-eclampsia pathophysiology.

Combined toxicity of imidacloprid and three insecticides to the earthworm, Eisenia fetida (Annelida, Oligochaeta).

Although the earthworm Eisenia fetida has been used in many ecotoxicological studies in recent years, most of these studies have only focused on assessing the effects of individual insecticides. In the present study, we aimed to compare the individual and combined toxic effects of imidacloprid and three insecticides (phoxim, chlorpyrifos, and lambda-cyhalothrin) on E. fetida. We showed that imidacloprid had the highest intrinsic toxicity to the worms in filter paper contact test, followed by phoxim and lambda-cyhalothrin, while the least toxicity was found from chlorpyrifos. Moreover, 14-day soil toxicity test revealed that the highest toxicity was still detected for imidacloprid with an LC50 value of 2.82 (2.61∼3.17) mg a.i. kg-1 dry weight (DW), followed by chlorpyrifos with an LC50 value of 384.9 (353.5∼440.3) mg a.i. kg-1 DW. Meanwhile, a relatively less toxicity was found for lambda-cyhalothrin with an LC50 value of 560.3 (475.9∼718.5) mg a.i. kg-1 DW, while the lowest toxicity to E. fetida was observed for phoxim with an LC50 value of 901.5 (821.3∼1017) mg a.i. kg-1 DW. In addition, significant synergistic responses were found from the ternary mixture of imidacloprid-phoxim-lambda-cyhalothrin and quaternary mixture of imidacloprid-phoxim-chlorpyrifos-lambda-cyhalothrin in both bioassay systems. Therefore, our findings highlighted that the simultaneous presence of several insecticides in the soil environment might lead to increased toxicity, resulting in serious damage to the nontarget organisms compared with individual insecticides.

Joint acute and endocrine disruptive toxicities of malathion, cypermethrin and prochloraz to embryo-larval zebrafish, Danio rerio.

It remains a daunting challenge to determine ecotoxicological risks of exposure to mixtures of endocrine disrupting chemicals (EDCs) in environmental toxicology. In the present study, we investigated acute and endocrine disruptive toxicities of cypermethrin (CPM), malathion (MAL), prochloraz (PRO) and their binary mixtures of MAL + CPM and MAL + PRO to the early life stages of zebrafish. In the acute lethal toxicity test, three pesticides exhibited different levels of toxicity to zebrafish larvae, and the order of toxicity was as follows: CPM > PRO > MAL. The binary mixture of MAL + CPM displayed a synergistic effect on zebrafish larvae after exposure for 24, 48, 72 and 96 h. However, binary mixture of MAL + PRO showed an antagonistic effect. To evaluate the estrogenic effect, the expression of genes in the hypothalamic-pituitary-gonadal axis was assessed after zebrafish embryos were exposed to CPM, MAL, PRO and their binary mixtures from blastula stage (1 h post-fertilization, 1 hpf) to 14 dpf (14 d post-fertilization). Our data indicated that the transcription patterns of many key genes (vtg1, vtg2, era, erß1, erß2, cyp19a1a and cyp19a1b) were affected in hatched zebrafish after exposure to CPM, MAL and PRO. Moreover, following exposure to binary mixtures of 1000 µg/L MAL +4 µg/L CPM and 1000 µg/L MAL +900 µg/L PRO, the gene expressions were significantly changed compared with the individual pesticides. Our data provided a better understanding of bidirectional interactions of toxic response induced by these pesticides.

Single and joint toxic effects of five selected pesticides on the early life stages of zebrafish (Denio rerio).

Instead of individual ones, pesticides are usually detected in water environment as mixtures of contaminants. Laboratory tests were conducted in order to investigate the effects of individual and joint pesticides (phoxim, atrazine, chlorpyrifos, butachlor and λ-cyhalothrin) on zebrafish (Denio rerio). Results from 96-h semi-static toxicity test indicated that λ-cyhalothrin had the greatest toxicity to the three life stages (embryonic, larval and juvenile stages) of D. rerio with LC50 values ranging from 0.0031 (0.0017-0.0042) to 0.38 (0.21-0.53) mg a.i. L-1, followed by butachlor and chlorpyrifos with LC50 values ranging from 0.45 (0.31-0.59) to 1.93 (1.37-3.55) and from 0.28 (0.13-0.38) to 13.03 (7.54-19.71) mg a.i. L-1, respectively. In contrast, atrazine showed the least toxicity with LC50 values ranging from 6.09 (3.34-8.35) to 34.19 (24.42-51.9) mg a.i. L-1. The larval stage of D. rerio was a vulnerable period to most of the selected pesticides in the multiple life stages tested. Pesticide mixtures containing phoxim and λ-cyhalothrin exerted synergistic effects on the larvae of D. rerio. Moreover, the binary mixture of phoxim-atrazine also displayed synergistic response to zebrafish. It has been assumed that most chemicals are additive in toxicity. Therefore, it is crucial to clarify the synergistic interaction for pesticide regulators and environment managers. In the present study, our data provided a clear picture on ecological risk of these pesticide mixtures to aquatic organisms. Moreover, joint effects play a more important role than individual ones, which require more attention when defining standard for water environment quality and risk assessment protocols.

Individual and mixture effects of five agricultural pesticides on zebrafish (Danio rerio) larvae.

In the present study, we evaluated the individual and mixture toxicities of imidacloprid and other four pesticides (atrazine, chlorpyrifos, butachlor, and λ-cyhalothrin) to the zebrafish (Danio rerio) larvae in order to clarify the interactive effects of pesticides on aquatic organisms. Results from the 96-h semi-static toxicity test indicated that chlorpyrifos, λ-cyhalothrin, and butachlor had the highest toxicities to D. rerio with an LC50 value ranging from 0.28 (0.13∼0.38) to 0.45 (0.31∼0.59) mg AI L-1, followed by atrazine with an LC50 value of 15.63 (10.71∼25.76) mg AI L-1, while imidacloprid exhibited the least toxicity to the organisms with an LC50 value of 143.7 (99.98∼221.6) mg AI L-1. Seven pesticide mixtures (two binary mixtures of imidacloprid + atrazine and imidacloprid + λ-cyhalothrin, two ternary mixtures of imidacloprid + atrazine + λ-cyhalothrin and imidacloprid + butachlor + λ-cyhalothrin, two quaternary mixtures of imidacloprid + atrazine + chlorpyrifos + λ-cyhalothrin and imidacloprid + chlorpyrifos + butachlor + λ-cyhalothrin, and one quinquenary mixture of imidacloprid + atrazine + chlorpyrifos + butachlor + λ-cyhalothrin) exhibited synergistic effects with equitoxic ratio and equivalent concentration on the zebrafish. Our results highlighted that the simultaneous presence of several pesticides in the aquatic environment might lead to increased toxicity, causing serious damage to the aquatic ecosystems compared with their individual toxicities. Therefore, the toxic effects of both individual pesticides and their mixtures should be incorporated into the environmental risk evaluation of pesticides.

Field investigations of dissipations and residues of cyazofamid in soil and tomato: risk assessment of human exposure to cyazofamid via tomato intake.

Cyazofamid, as a fungicide of the novel cyanoimidazole chemical class, has been widely used to control tomato late blight. Understanding of cyazofamid residues in environment and crops is an essential prerequisite for its risk assessment. In this study, field investigations in four typical tomato-producing areas were conducted to explore the dissipation kinetics and residues of cyazofamid and its primary metabolite 4-chloro-5-p-tolylimidazole-2-carbonitrile (CCIM) in soil and tomato. A robust method using QuEChERS coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for simultaneous analysis of cyazofamid and CCIM, with limits of quantification of 0.33 and 3.8 µg/kg, respectively. Field trials showed that the half-lives of cyazofamid were 3.6-6.9 days in soil and 12.2-18.3 days in tomato. The total residues of cyazofamid and CCIM in tomato collected at three time intervals were all below 0.5 mg/kg. Moreover, the potential risks of total residues via tomato intake to ten population subgroups were evaluated. We found that the risk quotient values were all generally low (0.13-1.3%), indicating that the recommended dose of cyazofamid on tomato will not result in a consumer exposure exceeding the toxicological reference value. Here, the results of field investigation provided important information for further understanding the behavior and risk of cyazofamid in the natural environment.

Development of chemical isotope labeling liquid chromatography mass spectrometry for silkworm hemolymph metabolomics.

Silkworm (Bombyx mori) is a very useful target insect for evaluation of endocrine disruptor chemicals (EDCs) due to mature breeding techniques, complete endocrine system and broad basic knowledge on developmental biology. Comparative metabolomics of silkworms with and without EDC exposure offers another dimension of studying EDCs. In this work, we report a workflow on metabolomic profiling of silkworm hemolymph based on high-performance chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS) and demonstrate its application in studying the metabolic changes associated with the pesticide dichlorodiphenyltrichloroethane (DDT) exposure in silkworm. Hemolymph samples were taken from mature silkworms after growing on diet that contained DDT at four different concentrations (1, 0.1, 0.01, 0.001 ppm) as well as on diet without DDT as controls. They were subjected to differential 12C-/13C-dansyl labeling of the amine/phenol submetabolome, LC-UV quantification of the total amount of labeled metabolites for sample normalization, and LC-MS detection and relative quantification of individual metabolites in comparative samples. The total concentration of labeled metabolites did not show any significant change between four DDT-treatment groups and one control group. Multivariate statistical analysis of the metabolome data set showed that there was a distinct metabolomic separation between the five groups. Out of the 2044 detected peak pairs, 338 and 1471 metabolites have been putatively identified against the HMDB database and the EML library, respectively. 65 metabolites were identified by the dansyl library searching based on the accurate mass and retention time. Among the 65 identified metabolites, 33 positive metabolites had changes of greater than 1.20-fold or less than 0.83-fold in one or more groups with p-value of smaller than 0.05. Several useful biomarkers including serine, methionine, tryptophan, asymmetric dimethylarginine, N-Methyl-D-aspartic and tyrosine were identified. The changes of these biomarkers were likely due to the disruption of the endocrine system of silkworm by DDT. This work illustrates that the method of CIL LC-MS is useful to generate quantitative submetabolome profiles from a small volume of silkworm hemolymph with much higher coverage than conventional LC-MS methods, thereby facilitating the discovery of potential metabolite biomarkers related to EDC or other chemical exposure.

Individual and combined toxic effects of herbicide atrazine and three insecticides on the earthworm, Eisenia fetida.

In the present study, we evaluated the individual and combined toxic effects of herbicide atrazine and three insecticides (chlorpyrifos, lambda-cyhalothrin and imidacloprid) on the earthworm, Eisenia fetida. Results from 48-h filter paper test indicated that imidacloprid had the highest toxicity to E. fetida with an LC50 of 0.05 (0.041-0.058) µg a.i. cm(-2), followed by lambda-cyhalothrin and atrazine with LC50 values ranging from 4.89 (3.52-6.38) to 4.93 (3.76-6.35) µg a.i. cm(-2), while chlorpyrifos had the least toxicity to the worms with an LC50 of 31.18 (16.22-52.85) µg a.i. cm(-2). Results from 14-days soil toxicity test showed a different pattern of toxicity except that imidacloprid was the most toxic even under the soil toxicity bioassay system. The acute toxicity of atrazine was significantly higher than that of chlorpyrifos. In contrast, lambda-cyhalothrin was the least toxic to the animals under the soil toxicity bioassay system. The binary mixture of atrazine-lambda-cyhalothrin and ternary mixture of atrazine-chlorpyrifos-lambda-cyhalothrin displayed a significant synergistic effect on the earthworms under the soil toxicity bioassay. Our findings would help regulatory authorities understand the complexity of effects from pesticide mixtures on non-target organisms and provide useful information of the interaction of various pesticide classes detected in natural environment.

Joint acute toxicity of the herbicide butachlor and three insecticides to the terrestrial earthworm, Eisenia fetida.

The herbicide butachlor and three insecticides phoxim, chlorpyrifos, and lambda-cyhalotrhin are widely used pesticides with different modes of action. As most previous laboratory bioassays for these pesticides have been conducted solely based on acute tests with a single compound, only limited information is available on the possible combined toxicity of these common chemicals to soil organisms. In this study, we evaluated their mixture toxicity on the terrestrial earthworm, Eisenia fetida, with binary, ternary, and quaternary mixtures. Two different types of bioassays were employed in our work, including a contact filter paper toxicity test and a soil toxicity test. Mixture toxicity effects were assessed using the additive index method. For all of the tested binary mixtures (butachlor-phoxim, butachlor-chlorpyrifos, and butachlor-lambda-cyhalothrin), significant synergistic interactions were observed after 14 days in the soil toxicity assay. However, greater additive toxicity was found after 48 h in the contact toxicity bioassay. Most of the ternary and quaternary mixtures exhibited significant synergistic effects on the worms in both bioassay systems. Our findings would be helpful in assessing the ecological risk of these pesticide mixtures to soil invertebrates. The observed synergistic interactions underline the necessity to review soil quality guidelines, which are likely underestimating the adverse combined effects of these compounds.

Pretilachlor has the potential to induce endocrine disruption, oxidative stress, apoptosis and immunotoxicity during zebrafish embryo development.

The objectives of the present study were to investigate the toxic effects of pretilachlor on zebrafish during its embryo development. The results demonstrated that the transcription of genes involved in the hypothalamic-pituitary-gonadal/thyroid (HPG/HPT) axis was increased after exposure to 50, 100, 200 µg/L pretilachlor for 96 h, the aromatase activity, vitellogenin (VTG) and thyroid hormones T3 and T4 levels in zebrafish were also up-regulated simultaneously. Pretilachlor exposure induced a noticeable increase in ROS level, increased the transcription and level of antioxidant proteins (e.g., CAT, SOD and GPX). Moreover, the up-regulation of P53, Mdm2, Bbc3 expression and Caspase3 and Caspase9 activities in the apoptosis pathway suggested pretilachlor might trigger cell apoptosis in zebrafish. In addition, the transcription of CXCL-C1C, IL-1ß and IL-8 related to the innate immunity was down-regulated after pretilachlor exposure. These data suggested that pretilachlor could simultaneously induce endocrine disruption, apoptosis, oxidative stress and immunotoxicity during zebrafish embryo development.