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.

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.

New Residue Analysis Method for Four Task-Specific Ionic Liquids in Water, Soil and Plants.

With the extensive application of task-specific ionic liquids (TSILs), their environmental impact has attracted increasing attention. However, no studies involving residue analyses of TSILs have been reported in the literature thus far. In the present study, residues of four TSILs ([C2NH2MIm]BF4, [HOEMIm]BF4, [HOEMIm]NO3, [MOEMIm]BF4) were analyzed by high-performance liquid chromatography-tandem mass spectrometry. The limit of detection of instrument was approximately 10-15 g. Residual TSILs were extracted from soil and plant samples by the accelerated solvent extraction method. In water, soil and plants, the coefficient of variation was 0.38%-4.43%, and the method detection limits of the four TSILs were lower than 1.40 ng g-1. These results meet the standards of residue analysis. The present study can provide an analysis method for studying TSIL residues and toxicity in the environment.

Ecotoxicity evaluation of azoxystrobin on Eisenia fetida in different soils.

Azoxystrobin, a widely used broad-spectrum strobilurin fungicide, may pose a potential threat in agricultural ecosystems. To assess the ecological risk of azoxystrobin in real soil environments, we performed a study on the toxic effects of azoxystrobin on earthworms (Eisenia fetida) in three different natural soils (fluvo-aquic soil, black soil and red clay soil) and an artificial soil. Acute toxicity of azoxystrobin was determined by filter paper test and soil test. Accordingly, exposure concentrations of chronic toxicity were set at 0, 0.1, 1.0 and 2.5 mg kg-1. For chronic toxicity test, reactive oxygen species, activity of antioxidant enzymes (superoxide dismutase, catalase and peroxidase), detoxifying enzyme (glutathione transferase), level of lipid peroxidation (malondialdehyde) and level of oxygen damage of DNA (8-hydroxydeoxyguanosine) in earthworms were determined on the 7th, 14th, 21st, 28th, 42nd and 56th days after treatment. Both acute and chronic toxic results showed azoxystrobin exhibit higher toxicity in natural soil than in artificial soil, indicating that traditional artificial soil testing method underestimate ecotoxicity of azoxystrobin in a real agricultural environment on the earthworm population. Combining with the analysis of soil physicochemical properties, the present experiment provided scientific guidance for rational application of azoxystrobin in agricultural production systems.

Influence of isolated bacterial strains on the in situ biodegradation of endosulfan and the reduction of endosulfan- contaminated soil toxicity.

The recently discovered endosulfan-degrading bacterial strains Pusillimonas sp. JW2 and Bordetella petrii NS were isolated from endosulfan-polluted water and soil environments. The optimal conditions for the growth and biodegradation activity of the strains JW2 and NS were studied in detail. In addition, the ability of the strains JW2 and NS to biodegrade endosulfan in soils during in situ bioremediation experiments was investigated. At a concentration of 2 mg of endosulfan per kilogram of soil, both JW2 and NS had positive effects on the degradation of endosulfan; JW2 degraded 100% and 91.5% of α- and ß-endosulfan, respectively, and NS degraded 95.1% and 90.3% of α- and ß-endosulfan, respectively. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of soil samples showed the successful colonization of JW2 and NS, and the toxicity of the soil decreased, as determined by single-cell gel electrophoresis (SCGE) assays of Eiseniafetida and micronucleus (MN) assays of Viciafaba root tip cells. Furthermore, the metabolic products of the bacterially degraded endosulfan from the in situ experiments were identified as endosulfan ether and lactone. This study provided potentially foundational backgrounds information for the remediation of endosulfan-contaminated soil.

Effects of the herbicide mesotrione on soil enzyme activity and microbial communities.

Mesotrione (2-[4-(methylsulfonyl)-2-nithobenzoyl]-1, 3-cyclohexanedione) is a selective triketone herbicide that has been widely used in corn production for the past 15 years. However, its potential for risk to soil ecosystems is poorly documented. The present study investigated the soil enzyme activity and soil microbial community responses to a 20 days' mesotrione exposure at doses of 0.1, 1.0 and 5.0 mg/kg. On days 2, 5, 10 and 20, activities of soil ß-glucosidase, urease and acid phosphatase, soil microbe abundances, soil microbial community structure and abundance of the AOA-amoA and AOB-amoA genes were measured. Results showed that activities of urease and acid phosphatase were relatively stable, with no difference found between the mesotrione-treated group and control at the end of exposure. But ß-glucosidase activity was reduced in the 5.0 mg/kg mesotrione treatment. In the 1.0 and 5.0 mg/kg mesotrione-treated soil, abundance of bacteria, fungi and actinomycetes all were reduced. In the 0.1 mg/kg mesotrione-treated soil, only fungi abundance was reduced by the end of the exposure. The analysis of terminal restriction fragment length polymorphism (T-RFLP) revealed soil microbial community structure could be affected by mesotrione at all experimental doses, and microbial diversity declined slightly after mesotrione exposure. Abundance of AOA-amoA and AOB-amoA genes were reduced markedly in 1.0 and 5.0 mg/kg mesotrione-treated soil. The present study suggests that mesotrione at higher doses might induce negative impacts on soil microbes, a finding which merits additional research and which should be accounted for when application of this herbicide is considered.

Effects of 1-Alkyl-3-Methylimidazolium Nitrate on Soil Physical and Chemical Properties and Microbial Biomass.

Ionic liquids (ILs), also called room temperature ILs, are widely applied in many fields on the basis of their unique physical and chemical properties. However, numerous ILs may be released into and gradually accumulate in the environment due to their extensive use and absolute solubility. The effects of 1-alkyl-3-methylimidazolium nitrate ([Cnmim]NO3, n = 4, 6, 8) on soil pH, conductivity, cation exchange capacity, microbial biomass carbon, and microbial biomass nitrogen were examined at the doses of 1, 10, and 100 mg/kg on days 10, 20, 30, and 40. The results demonstrated that the soil pH decreased and the conductivity increased with increasing IL doses. No significant differences were observed in the soil cation-exchange capacity. All three of the tested ILs decreased the soil microbial biomass carbon and nitrogen. Additionally, there were few differences among the ILs with different alkyl chain lengths on the tested indicators except for the microbial biomass nitrogen. The present study addressed a gap in the literature regarding the effects of the aforementioned ILs with different alkyl side chains on the physicochemical properties of soil, and the results could provide the basic data for future studies on their toxicity to soil organisms, such as earthworms and soil microbes.

Effects of 1-octyl-3-methylimidazolium nitrate on the microbes in brown soil.

The toxicity of ionic liquids (ILs) on soil organisms has aroused wide attention due to their high-solubility. The present investigation focused on the toxicity of 1-octyl-3-methylimidazolium nitrate ([C8mim]NO3) on the microbial populations (bacteria, fungi, and actinomycetes), soil enzyme (urease, dehydrogenase, acid phosphatase, and ß-glucosidase) activities, microbial community diversity using terminal restriction fragment length polymorphism (T-RFLP), and abundance of the ammonia monooxygenase (amoA) genes of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) using quantitative real-time polymerase chain reaction (q-PCR) in brown soil at each trial with doses of 0, 1.0, 5.0, and 10.0mg/kg on days 10, 20, 30, and 40. The contents of [C8mim]NO3 in soil were measured using high performance liquid chromatography with recoveries of 84.3% to 85.2%, and changed less than 10% during the experimental period. A significant decrease was observed from the bacteria, fungi and actinomycetes populations at 10.0mg/kg, at which the urease activity was inhibited and the ß-glucosidase activity was stimulated on days 20, 30, and 40. In addition, [C8mim]NO3 inhibited the dehydrogenase activity at 10mg/kg on days 30 and 40 and the acid phosphatase activity on day 20. The diversity of the soil microbial community and the gene abundance of AOA- and AOB- amoA were also inhibited. Furthermore, the present investigation provided more scientific information for the toxicity evaluation of ILs in soil.

Determination of Residual Concentration of Ionic Liquids with Different Anions and Alkyl-Chain Lengths in Water and Soil Samples.

Considering the wide synthesis and application of ionic liquids (ILs), the toxicity of ILs has recently gained growing attention. However, few studies focused on IL determination methods in environmental samples. In the present study, we implemented the determination methods for the 12 ILs with different chemical structures using high-performance liquid chromatography (HPLC) and ultraviolet (UV) spectrophotometry. The optimum conditions for extraction of ILs from soil samples were also obtained by single-factor experiments and response surface methodology (RSM). The instrument detection limits (IDLs) reached 10-10 g. Compared to the use of UV, HPLC had the standard curve with stronger correlation (R2 ≥ 0.999) and lower detection limit. We therefore used HPLC to detect the contents of ILs in water and soil samples. A standard adding method was used for the reliability test of the above methods. The average recovery in water samples was 90.46%-108.83%, and the coefficient of variation (CV) was 0.51%-9.07%. The method detection limits (MDLs) were below 0.1 mg/L. The optimized IL extraction conditions in soil samples were as follows: The ratio of methanol and saturated ammonium chloride was 90:10, the ultrasonic time was 50 min, and the power was 350 W. The average recovery in soil samples was 70.39%-85.30%, and the CV was 0.50%-9.99%. The MDLs were below 1 mg/kg. These results using the aforementioned methods met the standards of residue analysis. The present study can provide scientific analysis methods and a basis for evaluation of the study of IL residues in environmental samples.

Acute Toxicity of Imidazole Nitrate Ionic Liquids with Varying Chain Lengths to Earthworms (Eisenia foetida).

When ionic liquids (ILs) first came into use, we thought that they were safe. However, upon further investigation, researchers found that ILs are not harmless. In this study, the model soil organism, earthworms (Eisenia foetida), were used to study the acute toxicity of imidazole nitrate ionic liquids with varying chain lengths from 2 to 12. The experiment used two different methods, a filter paper contact test (48 h) and an artificial soil test (14 days), to determine the toxicity. These results demonstrated that the toxicity increased with the length of carbon chains until C8 and that the cut-off effect occurred at 1-octyl-3-methyl imidazole nitrates.Then, the toxicity began to increase again. At the same time, the concentrations of [C10mim]NO3 and [C12mim]NO3 were determined by high performance liquid chromatography and demonstrated that ILs were stable throughout the experiment. The present study revealed the acute toxicity of ILs with varying chain lengths.

Tissue-Specific Accumulation, Depuration, and Transformation of Triphenyl Phosphate (TPHP) in Adult Zebrafish (Danio rerio).

Understanding bioaccumulation and metabolism is critical for evaluating the fate and potential toxicity of compounds in vivo. We recently investigated, for the first time, the bioconcentration and tissue distribution of triphenyl phosphate (TPHP) and its main metabolites in selected tissues of adult zebrafish. To further confirm the metabolites, deuterated TPHP (d15-TPHP) was used in the exposure experiments at an environmentally relevant level (20 µg/L) and at 1/10 LC50 (100 µg/L). After 11-14 days of exposure to 100 µg/L of d15-TPHP, the accumulation and excretion of d15-TPHP reached equilibrium, at which point the intestine contained the highest d15-TPHP (µg/g wet weight, ww) concentration (3.12 ± 0.43), followed by the gills (2.76 ± 0.12) > brain (2.58 ± 0.19) > liver (2.30 ± 0.34) ≫ muscle (0.53 ± 0.04). The major metabolite of d15-TPHP, d10-diphenyl phosphate (d10-DPHP), was detected at significantly higher contents in the liver and intestine, at levels up to 3.0-3.5 times those of d15-TPHP. The metabolic pathways of TPHP were elucidated, including hydrolysis, hydroxylation, and glucuronic acid conjugation after hydroxylation. Finally, a physiologically based toxicokinetic (PBTK) model was used to explore the key factors influencing the bioaccumulation of d15-TPHP in zebrafish. These results provide important information for the understanding of the metabolism, disposition, and toxicology of TPHP in aquatic organisms.

Impairment of reproduction of adult zebrafish (Danio rerio) by binary mixtures of environmentally relevant concentrations of triclocarban and inorganic mercury.

Effects of chemical mixtures at environmentally relevant concentrations on endocrine systems of aquatic organisms are of concern. Triclocarban (TCC) and inorganic mercury (Hg2+) are ubiquitous in aquatic environments, and are known to interfere with endocrine pathways via different mechanisms of toxic action. However, effects of mixtures of the two pollutants on aquatic organisms and associated molecular mechanisms were unknown. This study examined effects of binary mixtures of TCC and Hg2+ on histopathological and biochemical alteration of reproductive organs in zebrafish (Danio rerio) after 21 d exposure. The results showed that: 1) At concentrations studied, TCC alone caused little effect on hepatic tissues, but it aggravated lesions in liver caused by Hg2+ via indirect mechanisms of disturbing homeostasis and altering concentrations of hormones; 2) Histological lesions were more severe in gonads of individuals, especially males, exposed to the binary mixture. Exposure to TCC alone (2.5 or 5µg/L) (measured concentration 140 or 310ng/L) or Hg2+ alone (5µg/L or 10µg/L (measured concentration 367 or 557ng/L) slightly retarded development of oocytes, whereas co-exposure to nominal concentrations of 5µg/L TCC and 10µg /L Hg2+ promoted maturation of oocytes. In males, maturation of sperm was slightly delayed by exposure to either TCC or Hg2+, while their combinations caused testes to be smaller and sperm to be fewer compared with fish exposed to either of the contaminants individually; 3) Lesions observed in fish exposed to binary mixtures might be due to altered transcription of genes involved in steroidogenesis, such as cyp19a, 3beta-HSD, cyp17, 17beta-HSD and modulated concentrations of testosterone and estradiol in blood plasma. The observed results further support the complexity of toxic responses of fish exposed to lesser concentrations of binary chemical mixtures. Since it is impossible to collect empirical information in controlled studies of all possible combinations of toxicants, the application of omics methods might improve the predictive capabilities of results of single classes of chemicals.

Oxidative stress and genotoxicity of the ionic liquid 1-octyl-3-methylimidazolium bromide in zebrafish (Danio rerio).

Ionic liquids (ILs) have a great reputation due to their negligible volatility, designability, good stability, and ability to be recycled. They are considered to be "green" solvents and have great promise in many fields. In recent years, the toxicities of ILs have garnered increasing attention as reported by a number of studies. However, previous studies have primarily focused on their lethal toxicities, and data were limited on their toxic effects at nonlethal doses. We performed a study on the toxic effects of 1-octyl-3-methylimidazolium bromide ([Omim]Br) on zebrafish. During a 28-day period, male and female zebrafish were separately exposed to sequential concentrations (0, 5, 10, 20, and 40 mg/L) of [Omim]Br. Fishes were sampled after 7, 14, 21, and 28 days of exposure, and reactive oxygen species (ROS) levels, activities of antioxidant enzymes (superoxide dismutase and catalase), lipid peroxidation (LPO), and DNA damage in fish livers were measured. ROS, LPO, and DNA damage were all induced by the ionic liquid, and antioxidant enzyme activities increased at the beginning and then decreased. These phenomena demonstrate that [Omim]Br can induce oxidative stress and DNA damage in zebrafish.

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.

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.

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.

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).

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.