Emerging disinfection byproducts 3-bromine carbazole induces cardiac developmental toxicity via aryl hydrocarbon receptor activation in zebrafish larvae.

3-bromine carbazole (3-BCZ) represents a group of emerging aromatic disinfection byproducts (DBP) detected in drinking water; however, limited information is available regarding its potential cardiotoxicity. To assess its impacts, zebrafish embryos were exposed to 0, 0.06, 0.14, 0.29, 0.58, 1.44 or 2.88 mg/L of 3-BCZ for 120 h post fertilization (hpf). Our results revealed that ≥1.44 mg/L 3-BCZ exposure induced a higher incidence of heart malformation and an elevated pericardial area in zebrafish larvae; it also decreased the number of cardiac muscle cells and thins the walls of the ventricle and atrium while increasing cardiac output and impeding cardiac looping. Furthermore, 3-BCZ exposure also exhibited significant effects on the transcriptional levels of genes related to both cardiac development (nkx2.5, vmhc, gata4, tbx5, tbx2b, bmp4, bmp10, and bmp2b) and cardiac function (cacna1ab, cacna1da, atp2a1l, atp1b2b, atp1a3b, and tnnc1a). Notably, N-acetyl-L-cysteine, a reactive oxygen species scavenger, may alleviate the failure of cardiac looping induced by 3-BCZ but not the associated cardiac dysfunction or malformation; conversely, the aryl hydrocarbon receptor agonist CH131229 can completely eliminate the cardiotoxicity caused by 3-BCZ. This study provides new evidence for potential risks associated with ingesting 3-BCZ as well as revealing underlying mechanisms responsible for its cardiotoxic effects on zebrafish embryos.

Effects of farmland residual mulch film-derived microplastics on the structure and function of soil and earthworm Metaphire guillelmi gut microbiota.

Microplastics derived from polyethylene (PE) mulch films are widely found in farmland soils and present considerable potential threats to agricultural soil ecosystems. However, the influence of microplastics derived from PE mulch films, especially those derived from farmland residual PE mulch films, on soil ecosystems remains unclear. In this study, we analyzed the bacterial communities attached to farmland residual transparent PE mulch film (FRMF) collected from peanut fields and the different ecological effects of unused PE mulch film-derived microplastics (MPs) and FRMF-derived microplastics (MPs-aged) on the soil and earthworm Metaphire guillelmi gut microbiota, functional traits, and co-occurrence patterns. The results showed that the assembly and functional patterns of the bacterial communities attached to the FRMF were clearly distinct from those in the surrounding farmland soil, and the FRMF enriched some potential plastic-degrading and pathogenic bacteria, such as Nocardioidaceae, Clostridiaceae, Micrococcaceae, and Mycobacteriaceae. MPs substantially influenced the assembly and functional traits of soil bacterial communities; however, they only significantly changed the functional traits of earthworm gut bacterial communities. MPs-aged considerably affected the assembly and functional traits of both soil and earthworm gut bacterial communities. Notably, MPs had a more remarkable effect on nitrogen-related functions than the MPs-aged in numbers for both soil and earthworm gut samples. Co-occurrence network analysis revealed that both MPs and MPs-aged enhanced the synergistic interactions among operational taxonomic units (OTUs) of the composition networks for all samples. For community functional networks, MPs and MPs-aged enhanced the antagonistic interactions for soil samples; however, they exhibited contrasting effects for earthworm gut samples, as MPs enhanced the synergistic interactions among the functional contents. These findings broaden and deepen our understanding of the effects of FRMF-derived microplastics on soil ecosystems, suggesting that the harmful effects of aged plastics on the ecological environment should be considered.

Combined toxic effects of polystyrene microplastics and 3,6-dibromocarbazole on zebrafish (Danio rerio) embryos.

Microplastics (MPs) and polyhalogenated carbazoles (PHCZs) are widely detected in the aquatic environment, and their ecological risks have become a research focus. Although there is an extensive co-distribution of MPs and PHCZs, their combined toxicity to aquatic organisms is still unclear. This study investigated the toxic effects of polystyrene microplastics (PS-MPs) and 3,6-dibromocarbazole (3,6-DBCZ) on zebrafish embryos by individual/combined exposure. This study showed that individual or combined exposure of PS-MPs (10 mg/L) and 3,6-DBCZ (0.5 mg/L) could significantly increase the rate of zebrafish embryo deformity, whereas no significant effect was observed on mortality and hatching rate. Furthermore, exposure to 3,6-DBCZ or PS-MPs increased reactive oxygen species (ROS) levels in zebrafish embryos, and the resulting oxidative stress induced apoptosis. Comparably, the levels of oxidative stress and apoptosis in zebrafish embryos were significantly reduced with the combined exposure of 3,6-DBCZ and PS-MPs. These observations suggest that the combined exposure of 3,6-DBCZ and PS-MPs has an antagonistic effect on oxidative stress and apoptosis. Fluorescence PS-MPs tracing and 3,6-DBCZ enrichment analysis showed that, with the protection of chorion, the entry of PS-MPs (5 and 50 µm) into the embryonic stage (55 hpf) of zebrafish was prevented. Moreover, after exposure for 96-144 hpf, PS-MPs served as a carrier to promote the 3,6-DBCZ accumulation and its dioxin-like toxicity in zebrafish larvae through ingestion. Compared with 5-µm PS-MPs, 50-µm PS-MPs promoted higher accumulation and dioxin-like toxicity of 3,6-DBCZ in zebrafish larvae. These findings provide that MPs can be used as an important carrier of PHCZs, influencing their toxicity and bioaccumulation in the organisms.

Effects of the polyhalogenated carbazoles 3-bromocarbazole and 1,3,6,8-tetrabromocarbazole on soil microbial communities.

Soil ecosystems are being more contaminated with polyhalogenated carbazoles (PHCZs), which raising much attention about their impact on soil microorganisms. 3-Bromocarbazole (3-BCZ) and 1,3,6,8-tetrabromocarbazole (1,3,6,8-TBCZ) are two typical PHCZs with high detection rates in the soil environment. However, ecological risk research on these two PHCZs in soil is still lacking. In the present study, after 80 days of exposure, the ecological influence of 3-BCZ and 1,3,6,8-TBCZ was investigated based on 16S rDNA sequencing, ITS sequencing, gene (16S rDNA, ITS, amoA, nifH, narG and cbbL) abundance and soil enzyme activity. The results showed that the bacterial 16S rDNA gene abundance significantly decreased under 3-BCZ and 1,3,6,8-TBCZ exposure after 80 days of incubation. The fungal ITS gene abundance significantly decreased under 1,3,6,8-TBCZ (10 mg/kg) exposure. PHCZs contributed to the alteration of bacteria and fungi community abundance. Bacteria Sphingomonas, RB41 and fungus Mortierella, Cercophora were identified as the most dominant genera. The two PHCZs consistently decreased the relative abundance of Sphingomonas, Lysobacter, Dokdonella, Mortierella and Cercophora etc at 80th day. These keystone taxa are related to the degradation of organic compounds, carbon metabolism, and nitrogen metabolism and may thus have influence on soil ecological functions. Bacterial and fungal functions were estimated using functional annotation of prokaryotic taxa (FAPROTAX) and fungi functional guild (FUNGuild), respectively. The nitrogen and carbon metabolism pathway were affected by 3-BCZ and 1,3,6,8-TBCZ. The soil nitrogen-related functions of aerobic ammonia oxidation were decreased but the soil carbon-related functions of methanol oxidation, fermentation, and hydrocarbon degradation were increased at 80th day. The effects of 3-BCZ and 1,3,6,8-TBCZ on the abundances of the amoA, nifH, narG, and cbbL genes showed a negative trend. These results elucidate the ecological effects of PHCZs and extend our knowledge on the structure and function of soil microorganisms in PHCZ-contaminated ecosystems.

Effect of flumetsulam alone and coexistence with polyethylene microplastics on soil microbial carbon and nitrogen cycles: Elucidation of bacterial community structure, functional gene expression, and enzyme activity.

Flumetsulam (FLU) is a new class of broad-spectrum herbicides. With the widespread use of plastic products, polyethylene (PE) microplastics (MPs) may remain in the soil. It is possible for these two novel contaminants to co-exist in the soil environment. In the present study, we used brown soil as the test soil and determined the toxicity of FLU at 0.05, 0.5 and 2.5 mg kg-1 alone and in combination with PE MPs (1%) on soil microorganisms. The obtained results demonstrated that the exposure of FLU and FLU+MPs had an inhibitory effect on the numbers of bacteria and fungi. In addition, FLU and FLU+MPs caused changes in the relevant functional bacterial genera, favored nitrogen fixation and denitrification, and promoted soil carbon fixation, but inhibited nitrification. Compared to FLU exposure alone, exposure to FLU+MPs gave rise to significant differences in soil bacterial community composition, but did not affect carbon and nitrogen cycling. The integrated biomarker response results indicated that the toxicity of FLU and FLU+MPs to soil microorganisms increased with increasing concentrations of FLU. The present experiment clarified the toxicological effects of co-exposure of FLU and MPs on microorganisms and filled the toxicological data gap of FLU.

Are PFBS, PFHxS, and 6:2FTSA more friendly to the soil environment compared to PFOS? A new insight based on ecotoxicity study in soil invertebrates (Eisenia fetida).

As alternatives to perfluorooctane sulfonate (PFOS) with shorter carbon chains or lower proportion of fluorine atoms, perfluorobutane sulfonate (PFBS), perfluorohexane sulfonate (PFHxS), and 6:2 fluorotelomer sulfonic acid (6:2FTSA) have been detected in various environmental media. However, it is unclear whether the toxicity of these alternatives is lower than that of PFOS. Therefore, this study investigated the toxicity and differences in PFBS, PFHxS, 6:2FTSA, and PFOS (0.2 mg/kg) after 56 d of exposure using the common invertebrate Eisenia fetida in soil as the test organism. The results showed that although PFOS, PFBS, PFHxS, and 6:2FTSA induced oxidative stress and apoptosis in earthworms and led to developmental and reproductive toxicity in terms of comprehensive toxicity, PFHxS > PFOS > PFBS >6:2FTSA. To reveal the mechanisms underlying the differences in toxicity between the alternatives and PFOS, we conducted molecular docking and transcriptomic analyses. The results indicated that, unlike PFOS, PFBS, and PFHxS, 6:2FTSA did not cause significant changes in antioxidant enzyme activity at the molecular level. Furthermore, PFOS exposure caused disorder in the nervous and metabolic systems of earthworms, and PFHxS disrupted energy balance and triggered inflammatory responses, which may be important reasons for the higher toxicity of these compounds. In contrast, exposure to 6:2FTSA did not result in adverse transcriptomic effects, suggesting that 6:2FTSA exerted the least molecular-scale toxicity in earthworms. The results of this study provide new insights into the environmental safety of using PFBS, PFHxS, and 6:2FTSA as alternatives to PFOS.

Analysis of the toxic mechanisms of fluoxastrobin on the earthworm (Eisenia fetida) using transcriptomics.

Fluoxastrobin (FLUO), one of the best-selling strobilurin fungicides, could prevent fungal diseases from oilseed crops, fruits, grains, and vegetables. The widespread use of FLUO leads to the continuous accumulation of FLUO in soil. Our previous studies have demonstrated that FLUO exhibited different toxicity in artificial soil and three natural soils (fluvo-aquic soils, black soils, and red clay). The toxicity of FLUO was greater in natural soil than the artificial soil, specifically, showed the highest toxicity in fluvo-aquic soils. To better investigate the mechanism of FLUO toxicity to earthworms (Eisenia fetida), we selected fluvo-aquic soils as representative soil and used transcriptomics to study the gene expression in earthworms after FLUO exposure. The results demonstrated that the differentially expressed genes in earthworms after FLUO exposure mainly presented in pathways involving protein folding, immunity, signal transduction, and cell growth. It may be the reason why FLUO exposure stressed the earthworms and affected their normal growth activities. The present study fills gaps in the literature regarding the soil bio-toxicity of strobilurin fungicides. It also sounds the alarm for the application of such fungicides even at the low concentration (0.1 mg kg-1).

Effects of the fungicide trifloxystrobin on the structure and function of soil bacterial community.

Trifloxystrobin has been widely applied to prevent fungal diseases because of its high efficiency and desirable safety characteristics. In the present study, the effects of trifloxystrobin on soil microorganisms were integrally investigated. The results showed that trifloxystrobin inhibited urease activity, promoted dehydrogenase activity. Downregulated expressions of the nitrifying gene (amoA), denitrifying genes (nirK and nirS), and carbon fixation gene (cbbL) were also observed. Soil bacterial community structure analysis showed that trifloxystrobin changed the abundance of bacteria genera related to nitrogen and carbon cycle in soil. Through the comprehensive analysis of soil enzymes, functional gene abundance, and soil bacterial community structure, we concluded that trifloxystrobin inhibited both nitrification and denitrification of soil microorganisms, and also diminished the carbon-sequestration ability. Integrated biomarker response analysis showed that dehydrogenase and nifH were the most sensitive indicators of trifloxystrobin exposure. It provides new insights about trifloxystrobin environmental pollution and its influence on soil ecosystem.

The effect of TiO2NPs on cloransulam-methyl toxicity to earthworm (Eisenia fetida).

Cloransulam-methyl is a new herbicide and has broad application prospect. However, the effect of cloransulam-methyl on earthworm have yet to be clarified. As more and more titanium dioxide nanoparticles (TiO2NPs) enter the soil, cloransulam-methyl and TiO2NPs have a risk of co-exposure, but the effect of TiO2NPs on cloransulam-methyl toxicity is unknown. In the study, the ecotoxicity of cloransulam-methyl (0.1, 1 mg kg-1) on earthworm and the effect of TiO2NPs (10 mg kg-1) on cloransulam-methyl toxicity was investigated after exposure for 28 and 56 d. Exposure tests showed cloransulam-methyl and cloransulam-methyl + TiO2NPs promoted the accumulation of reactive oxygen species, malondialdehyde and 8-hydroxydeoxyguanosine, increased the activities of superoxide dismutase and catalase, resulted in lipid peroxidation and DNA damage. Besides, the results at the genetic level showed cloransulam-methyl and cloransulam-methyl + TiO2NPs altered the expression of physiologically-related genes, which demonstrated that cloransulam-methyl and cloransulam-methyl + TiO2NPs induced oxidative stress and cell apoptosis, and disturbed the normal reproduction in earthworm. The results of comprehensive toxicity comparison indicated cloransulam-methyl and TiO2NPs co-exposure has higher toxicity compared to cloransulam single exposure. Our results suggest that TiO2NPs can enhance the toxicity of cloransulam-methyl on Eisenia fetida in terms of oxidative stress, cell apoptosis and reproduction aspects. Based on above studies, it is of great importance for evaluating the risk of cloransulam-methyl co-exposure with TiO2NPs in soil.

Development toxicity and cytotoxicity of pyroxsulam on embryos and adults of zebrafish (Danio rerio).

Pyroxsulam is a typical triazolopyrimidine sulfonamide herbicide, which plays a vital role in weed control for wheat production. Previous studies on pyroxsulam have mainly focused on weed resistance, control effects, and soil microorganisms, however, research on aquatic ecotoxicity is lacking. This study evaluated the toxicity of pyroxsulam in zebrafish embryos (120 h) and adults (14 and 28 d). Chronic exposure to pyroxsulam (0.2, 2, and 10 µg L-1) reduced reactive oxygen species (ROS) content in embryos but increased ROS content in adults. Pyroxsulam caused lipid peroxidation and DNA damage in embryos and adults. The expression of p53, bcl-2/bax, and caspase 3/8/9 indicated that pyroxsulam inhibited apoptosis in embryos but induced apoptosis in adults. By calculating integrated biomarker response, apoptosis was more readily affected than oxidative damage in embryos and adults. The toxicity of pyroxsulam increased with increasing concentration, however, with increasing exposure time, the toxicity of pyroxsulam to adults decreased. Pyroxsulam exerted toxic effects on zebrafish at different life stages (embryos and adults), and different stages had different toxicity. These results indicate that in future studies on the toxicity of pyroxsulam to zebrafish, different life stages (embryos and adults) need to be studied. The present study evaluated the toxicity of environmentally relevant concentrations of pyroxsulam to zebrafish embryos and adults, providing worthy data for assessing its effects on aquatic ecosystems.

Characterization of the responses of soil micro-organisms to azoxystrobin and the residue dynamics of azoxystrobin in wheat-corn rotation fields over two years.

Azoxystrobin, a high-efficiency and broad-spectrum strobilurin fungicide, has been widely used in global agricultural production. However, the effects of azoxystrobin on soil micro-organisms have scarcely been studied, and relevant experiments are usually conducted under laboratory conditions using active ingredient. Therefore, the effects of azoxystrobin on soil micro-organisms when applied to actual farmland are unknown. We sought to address this knowledge gap in this study, where we studied the effects of azoxystrobin on soil micro-organisms in a wheat-corn rotation field over two years. The results indicate that after two years of azoxystrobin application the activities of soil enzymes were inhibited, and the abundance of functional genes related to the nitrogen and carbon cycle were inhibited, which change the abundance of soil microbial bacteria of genera. As a consequence, the soil nitrogen and carbon cycles were disturbed. In addition, azoxystrobin inhibited the abundance of functional bacteria related to organic pollutant degradation and soil metabolism, where the rate of azoxystrobin degradation diminished over time. Moreover, azoxystrobin significantly inhibited the soil-culturable microbial population. The integrated biomarker response (IBR) indicated that the soil-culturable microbial population can be used as a sensitive indicator of the effect of azoxystrobin on soil micro-organisms. The final levels of azoxystrobin residues measured in grains were less than 0.004 mg/kg, lower than the maximum residue limits in European Union and China. The results of this study provide a basis for suggestions regarding the appropriate use of azoxystrobin in addition to support for elucidating the interaction between biological macromolecules and pollutants.

New insights into the effects of chlorpyrifos on soil microbes: Carbon and nitrogen cycle related microbes in wheat/maize rotation agricultural field.

Chlorpyrifos, a broad-spectrum organophosphorus insecticide, has been widely detected worldwide and is a potential neurotoxin and endocrine disruptor. Besides, chlorpyrifos has been proven that have a negative effect on soil microbes. In the present study, chlorpyrifos formulation (LORSBAN®, 45% emulsifiable concentrate) was applied in an agricultural field at the recommended dose (R dose, 270.0 and 337.5 g a.i. ha-1 for wheat and maize respectively) and double recommended (DR) dose. Chlorpyrifos residue level and effect on soil microbes related to soil carbon and nitrogen cycle function were analyzed. Results showed that the half-lives of chlorpyrifos in wheat and maize field soil were 7.23-8.23 and 1.45-1.77 d, respectively. Application of chlorpyrifos at even DR dose did not result in unacceptable residual chlorpyrifos, where the final residual chlorpyrifos in wheat/maize (leaf, stem, and grain) was meet the requirement of the maximum residual limit (0.5 mg kg-1 for wheat and 0.05 mg kg-1 for maize) in China. Chlorpyrifos enhanced the activity of ß-glucosidase by increasing the relative abundance of Sphingosinicella and promoted the carbon cycle in wheat field. The changes of cbbLR and cbbLG gene abundance also confirmed that chlorpyrifos could affect the import and export of soil carbon pool. The effect of chlorpyrifos on soil N cycle was determined by changes in the abundance of the bacterial genus Gemmatimonas, which is associated with denitrification. Further analysis of N-cycle functional genes and urease activity showed that chlorpyrifos inhibited nitrogen fixation in wheat field, but promoted nitrogen fixation in maize field. In general, bacterial abundance, urease, and AOA-amoA gene could be early warning markers of chlorpyrifos contamination. The results demonstrated the negative effects of chlorpyrifos on soil microbes especially on soil C and N cycle in actual agricultural field. It provides new insights about chlorpyrifos environmental pollution and its effect on soil ecosystems.

Effect of florasulam on oxidative damage and apoptosis in larvae and adult zebrafish (Danio rerio).

Florasulam is widely used for weed control in wheat fields due to its high activity and low dosage. Previous studies on florasulam have focused on soil microbial and residue determination, however, its ecotoxicity to aquatic organisms is unclear. The toxicity of florasulam was evaluated in larvae (120 h) and adult (14 and 28 d) zebrafish. After florasulam (0.1 and 1 µg L-1) exposure, reactive oxygen species levels in larvae and adult zebrafish significantly increased and antioxidant system was activated. Florasulam induced lipid peroxidation in larvae and adult zebrafish. Florasulam did not cause DNA damage to larvae but caused DNA damage to adult zebrafish. Changes in caspase 3/8/9 genes indicated that apoptosis was induced in larvae but inhibited in adult zebrafish. By calculating integrated biomarker response, caspase 3 and malondialdehyde could be used as early warning indicators of florasulam effect on larvae and adult zebrafish, respectively. The higher the exposure concentration, the greater the toxicity of florasulam to larvae and adult zebrafish. Increasing exposure time resulted in higher toxicity to adult zebrafish. Florasulam has different toxicity at larvae and adult zebrafish. In future studies to investigate florasulam toxicity to zebrafish, different zebrafish life stages (larvae and adult) need to be studied.

Reproductive toxicity of enrofloxacin in Caenorhabditis elegans involves oxidative stress-induced cell apoptosis.

Fluoroquinolone antibiotics (FQs) that persist and bioaccumulate in the environment have aroused people's great concern. Here, we studied the adverse effects of FQs in soil animals of Caenorhabditis elegans via food-chronically exposure. The result shows C. elegans exposed to FQs exhibited reproductive toxicity with small-brood size and low-egg hatchability. To study the underlying mechanism, we conduct a deep investigation of enrofloxacin (ENR), one of the most frequently detected FQs, on nematodes which is one of commonly used animal indicator of soil sustainability. The concentration-effect curves simulated by the Hill model showed that the half effect concentrations (EC50) of ENR were (494.3 ± 272.9) µmol/kg and (107.4 ± 30.9) µmol/kg for the brood size and the hatchability, respectively. Differential gene expression between the control and the ENR-exposure group enriched with the oxidative stress and cell apoptosis pathways. The results together with the enzyme activity in oxidative stress and the cell corpses suggested that ENR-induced reproductive toxicity was related to germ cell apoptosis under oxidative stress. The risk quotients of some soil and livestock samples were calculated based on the threshold value of EC10 for the egg hatchability (2.65 µmol/kg). The results indicated that there was possible reproductive toxicity on the nematodes in certain agricultural soils for the FQs. This study suggested that chronic exposure to FQs at certain levels in environment would induce reproductive toxicity to the nematodes and might reduce the soil sustainability, alarming the environment risks of antibiotics abuse.

Toxicity comparison of atrazine on Eisenia fetida in artificial soil and three natural soils.

Atrazine has been widely used in the world and caused environmental pollution, especially soil pollution. When assessing the toxicity of atrazine in soil, most studies used standardized artificial soils, while few studies focused on the real soil environments. In the present study, three natural soils and artificial soil were selected as test soils to study and compare the toxicities of atrazine to Eisenia fetida. Acute toxicity of atrazine was determined by filter paper and soil tests. In chronic toxicity study, after atrazine exposure, the content of reactive oxygen species in Eisenia fetida significantly increased and showed a dose-response relationship. The activity changes of three antioxidant enzymes and glutathione transferase showed that atrazine had obvious oxidative stress effect on earthworms. The contents of malondialdehyde and 8-hydroxy deoxyguanosine in 0.1 and 1 mg/kg atrazine treatment groups were significantly higher than the control, indicating that medium and high concentrations of atrazine could cause lipid and DNA damage in Eisenia fetida. The acute toxicity results and the integrated biomarker response index for chronic toxicity indicated that the toxicity order of atrazine was: red clay > fluvo-aquic soil > artificial soil > black soil, and that the toxicity of atrazine in artificial soil was not representative of its toxicity in real soil environment. The results of correlation analysis showed that three soil property parameters of organic carbon, organic matter and sand were most related to the toxicity of atrazine.

Effects of ecotoxicity of penoxsulam single and co-exposure with AgNPs on Eisenia fetida.

Penoxsulam (PNX) and silver nanoparticles (AgNPs) are likely to coexist in soils due to continuous use. However, the ecotoxicity of PNX in earthworms and the effect of AgNPs on PNX toxicity are unknown. Therefore, the toxicity of PNX (0.05, 0.5, and 2.5 mg/kg) single and co-exposure with AgNPs (10 mg/kg) after 28 and 56 days on Eisenia fetida (E. fetida) was investigated from biochemical, genetic, histopathological, and transcriptomic aspects. The results showed that the low concentration of PNX (0.05PNX) had almost no effect on the biochemical level of E. fetida. However, the addition of AgNPs resulted in 0.05PNX causing E. fetida to produce excessive reactive oxygen species, and the activity of antioxidant and detoxification enzymes were interfered, resulting in lipid peroxidation and DNA damage. From the genetic level, even the lowest concentration of PNX can significantly interfere with the expression of functional genes, thus inducing oxidative stress and apoptosis and inhibited reproductive behavior in E. fetida. The integrated biomarker response results at the biochemical and genetic levels showed that the comprehensive toxicity of PNX and PNX + AgNPs on E. fetida was PNX dose-dependent. And the toxicity of all co-exposure groups was greater than that of the PNX only exposure groups. Furthermore, the addition of AgNPs significantly increased the damage of PNX on E. fetida intestinal tissue. Meanwhile, transcriptomic analysis showed that PNX + AgNPs had a greater effect on E. fetida than PNX single, and multiple pathways related to oxidative stress, inflammation, and cellular process regulation were disturbed. These results provide a basis for comprehensive evaluation of the ecotoxicity of PNX and confirm that the AgNPs does increase the ecotoxicity of PNX in soil environment.

Toxicity evaluation of chlorpyrifos and its main metabolite 3,5,6-trichloro-2-pyridinol (TCP) to Eisenia fetida in different soils.

The present study utilized a biomarker response method to evaluate the effect of 3,5,6-trichloro-2-pyridinol (TCP) in artificial and natural soils on Eisenia fetida after 7, 14, 28, 42 and 56 days exposure. Results indicated that TCP induced excessive reactive oxygen species, caused oxidative stress and DNA damage to Eisenia fetida. Biomarker responses were standardized to calculate the Integrated Biomarker Response (IBR) index. The IBR index of three enzymes (superoxide dismutase, catalase and glutathione S-transferase) activities showed that TCP induced the oxidative stress to E. fetida in red clay was stronger than in the other three soils. Specifically, chlorpyrifos exposure group showed a lower toxicity than TCP exposure group after 28 days exposure but a higher toxicity than TCP exposure group after 56 days exposure. Despite the deficiencies of this study, the above information is of great significance for assessing the risk of chlorpyrifos and its metabolite TCP pollution in soil ecosystems.

Polyhalogenated carbazoles (PHCZs) induce cardiotoxicity and behavioral changes in zebrafish at early developmental stages.

Polyhalogenated carbazoles (PHCZs) are widely present in the environment, and their health risks are of increasing concern. Available studies primarily confirm their dioxin-like toxicity mechanism based on biomarkers, such as aryl hydrocarbon receptor (AHR) and CYP1A1, while few studies have investigated their actual toxic effects at the level of individual organisms. In the present study, the developmental toxicity of two typical PHCZs with a high detection rate and high concentration in the environment (3,6-dichlorocarbazol (3,6-DCCZ) and 3,6-dibromocarbazole (3,6-DBCZ)) was investigated based on a fish embryo acute toxicity test (FET, zebrafish) and transcriptomics analysis. The 96 h LC50 values of 3,6-DCCZ and 3,6-DBCZ were 0.636 mg/L and 1.167 mg/L, respectively. Both tested PHCZs reduced the zebrafish heart rate and blocked heart looping at concentrations of 0.5 mg/L or higher. The swimming/escaping behavior of zebrafish larvae was more vulnerable to 3,6-DBCZ than 3,6-DCCZ. Transcriptomics assays showed that multiple pathways linked to organ development, immunization, metabolism and protein synthesis were disturbed in PHCZ-exposed fish, which might be the internal mechanism of the adverse effects. The present study provides evidence that PHCZs cause cardiac developmental toxicity and behavioral changes and improves our understanding of their health risks.

Toxicity evaluation of pyraclostrobin exposure in farmland soils and co-exposure with nZnO to Eisenia fetida.

Although the toxicity of pyraclostrobin (PYRA) to earthworms in artificial soil is well known, the toxicity of PYRA in farmland soils is yet to be explored in detail. Additionally, with more zinc oxide nanoparticles (nZnO) entering the soil environment, the risk of PYRA co-exposure with nZnO is increasing alarmingly. However, toxicity caused by this co-exposure of PYRA and nZnO is still unknown. Therefore, we assessed the biomarkers responses to reveal the toxicity of PYRA (0.1, 1, 2.5 mg/kg) on earthworms in farmland soils (black soil, fluvo-aquic soil, and red clay) and evaluated the biomarkers responses of Eisenia fetida exposed to PYRA (0.5 mg/kg)/PYRA+nZnO (10 mg/kg). Moreover, transcriptomic analysis was performed on E. fetida exposed to PYRA/PYRA+nZnO for 28 days to reveal the mechanism of genotoxicity. The Integrated Biomarker Responses (IBR) showed PYRA induced more severe oxidative stress and damage to E. fetida in farmland soils than that in artificial soil. The oxidative stress and damage induced by PYRA+nZnO were greater than that induced by PYRA. Transcriptomic analysis showed that PYRA and PYRA+nZnO significantly altered gene expression of both biological processes and molecular functions. These results provided toxicological data for PYRA exposure in three typical farmland soils and co-exposure with nZnO.

Comparison of the toxic effects of non-task-specific and task-specific ionic liquids on zebrafish.

Ionic liquids (ILs) are composed of only anions and cations and are liquid solvents at room temperature. Different functional groups were introduced into the ILs, conferring them with specific functions or purposes and thus forming special ILs, namely task-specific ILs (TSILs). Imidazolium-based ILs are the most widely used ILs in industrial production. To date, there have been some studies on the toxic effects of ILs on different organisms. However, the effect of functionalized groups on the toxicity of ILs is still unclear. In the present study, zebrafish were used as model organisms to study the toxic effects of 1-ethyl-3-methylimidazolium nitrate ([C2mim]NO3) and 1-hydroxyethyl-3-methylimidazolium nitrate ([HOC2mim]NO3). The results showed that both promoted an increase in reactive oxygen species (ROS) contents, leading to lipid peroxidation and DNA damage. Furthermore, integrated biological response analysis showed that [HOC2mim]NO3 was less toxic to zebrafish than [C2mim]NO3, which indicated that adding functional groups decreased the toxicity of ILs to organisms. The influence of chemical structure on IL toxicity was also reported. These results could provide a scientific basis for better synthesis and utilization of ILs in the future.