Method development, validation, and risk assessment of multiple pesticide residues of fruits in China.

In this study, a muti-residue analysis method of 40 pesticides in five different categories of fruits in China was developed based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Five hundred real samples were analyzed and assessed for the dietary exposure risk. The sample treatment method was optimized by comparing four clean-up methods. The matrix effects of different fruits were evaluated. The analytical method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, and precision. The results showed that the optimal method was the treatment by clean up with ODS (octadecylsilane) and MgSO4. The matrix effect was the strongest in orange and weakest in apple. The LOD and LOQ of pesticides were 0.04-5.9 µg kg-1 and 0.13-19.5 µg kg-1, respectively. The recoveries at three spiked levels were ranged from 71.2 to 115.2% with the RSDs from 0.1 to 19.6%. Twenty-two pesticides were detected in 500 fruit samples from the major production regions of China, with concentrations ranging from 0.1 to 1930 µg kg-1. A total of 13, 10, 9, 8, and 4 pesticides were detected in peach, orange, grape, apple, and strawberry. Both the acceptable daily intake (ADI) and acute reference dose (ARfD) for all the detected pesticides were lower than 100%, indicating that the dietary intake risks are acceptable and would not pose potential health risks.

Insights into spirotetramat-induced thyroid disruption during zebrafish (Danio rerio) larval development: An integrated approach with in vivo, in vitro, and in silico analyses.

Spirotetramat (SPT), a tetronic acid-derived insecticide, is implicated in reproductive and lipid metabolism disorders, as well as developmental toxicity in fish. While these effects are documented, the precise mechanisms underlying its developmental toxicity are not fully elucidated. In this study, zebrafish embryos (2 h post-fertilization, hpf) were exposed to four concentrations of SPT (0, 60, 120, and 240 µg/L) until 21 dpf (days post-fertilization). We delved into the mechanisms by examining its potential disruption of the thyroid endocrine system, employing in vivo, in vitro, and in silico assays. The findings showed notable developmental disturbances, including reduced hatching rates, shortened body lengths, and decelerated heart rates. Additionally, there was an increase in malformations and a decline in locomotor activity. Detailed analyses revealed that SPT exposure led to elevated thyroid hormone levels, perturbed the hypothalamic-pituitary-thyroid (HPT) axis transcript levels, amplified deiodinase type I (Dio1) and deiodinase type II (Dio2) activities, and both transcriptionally and proteomically upregulated thyroid receptor beta (TRß) in larvae. Techniques like molecular docking and surface plasmon resonance (SPR) confirmed SPT's affinity for TRß, consistent with in vitro findings suggesting its antagonistic effect on the T3-TR complex. These insights emphasize the need for caution in using tetronic acid-derived insecticides.

Core bacteria carrying the genes associated with the degradation of atrazine in different soils.

Atrazine residues can pose serious threats to soil ecology and human health. Currently, the underlying relationship between soil microbial communities and the degradation genes associated with atrazine degradation remains unclear. In this study, the degradation characteristics of atrazine was investigated in ten different soil types. Further, diversity and abundance of degradation genes and succession of the bacterial community were also studied. The degradation of 10 mg/kg atrazine in different soil types exhibited an initial rapid trend followed by a gradual slowdown, adhering to the first-order kinetic equation. Atrazine significantly increased the absolute abundance of atz degradation genes. The increase in the absolute abundance of atzC gene was the largest, whereas that of atzA gene was the smallest, and the trzD gene was only detected in the Binzhou loam soil. Co-occurrence network analysis showed that the number of potential bacterial hosts of atzC was the highest compared with the other atz genes. Atrazine also altered the structural composition of the soil microbial community. The relative abundances of Ochrobactrum, Nocardiopsis, Lactobacillus, and Brevibacterium was increased in the atrazine-treated soils, while those of Conexibate, Solirubacter, and Micromonospora was decreased significantly compared with the control. Additionally, four atrazine-degrading bacterial strains Rhizobium AT1, Stenotrophomonas AT2, Brevibacterium AT3, and Bacillus AT4 were isolated from the atrazine-treated soils. After 14 d for inoculation, their degradation rate for 10 mg/L atrazine ranged from 17.56 % to 30.55 %. Moreover, the relative abundances of the bacterial genera, including these four isolates, in the atrazine-treated soil were significantly higher than those in the control, indicating that they were involved in the synergistic degradation of atrazine in the soil. This study revealed the degradation characteristics of atrazine, distribution of degradation genes, and succession of microbial communities, and explored the internal relationship between microbial community structure and atrazine degradation mechanisms in different soil types.

Deciphering the degradation characteristics of the fungicides imazalil and penflufen and their effects on soil bacterial community composition, assembly, and functional profiles.

The adsorption-desorption and degradation characteristics of two widely applied fungicides, imazalil and penflufen, and the responses of soil bacterial diversity, structure, function, and interaction after long-term exposure were systemically studied in eight different soils. The adsorption ability of imazalil in soil was significantly higher than that of penflufen. Both imazalil and penflufen degraded slowly in most soils following the order: imazalil > penflufen, with soil pH, silt, and clay content being the potential major influencing factors. Both imazalil and penflufen obviously inhibited the soil microbial functional diversity, altered the soil bacterial community and decreased its diversity. Although exposure to low and high concentrations of imazalil and penflufen strengthened the interactions among the soil bacterial communities, the functional diversity of the co-occurrence network tended to be simple at high concentrations, especially in penflufen treatment. Both imazalil and penflufen markedly disturbed soil nitrogen cycling, especially penflufen seriously inhibited most nitrogen cycling processes, such as nitrogen fixation and nitrification. Meanwhile, sixteen and ten potential degradative bacteria of imazalil and penflufen, respectively, were found in soils, including Kaistobacter and Lysobacter. Collectively, the long-term application of imazalil and penflufen could cause residual accumulation in soils and subsequently result in serious negative effects on soil ecology.

Earthworms synergize with indigenous soil functional microorganisms to accelerate the preferential degradation of the highly toxic S-enantiomer of the fungicide imazalil in soil.

The roles of soil and earthworm gut microorganisms in the degradation of the chiral fungicide imazalil (IMA) enantiomers were systemically studied in soil-earthworm systems. S-IMA degraded slower than R-IMA in soil without earthworms. After the addition of earthworms, S-IMA degraded faster than R-IMA. Methylibium was the potential degradative bacterium likely related to the preferential degradation of R-IMA in soil. However, the addition of earthworms significantly decreased the relative abundance of Methylibium, especially in R-IMA-treated soil. Meanwhile, a new potential degradative bacterium Aeromonas first appeared in soil-earthworm systems. Compared with enantiomer-treated soil, the relative abundance of indigenous soil bacterium Kaistobacter significantly boomed in enantiomer-treated soil with earthworms. Interestingly, Kaistobacter in the earthworm gut also obviously increased after exposure to enantiomers, particularly in S-IMA-treated soil, which was associated with the significant increase in Kaistobacter in soil. More importantly, the relative abundances of Aeromonas and Kaistobacter in S-IMA-treated soil were obviously higher than those in R-IMA-treated soil after the addition of earthworms. Moreover, these two potential degradative bacteria were also potential bacterial hosts of the biodegradation genes p450 and bph. Collectively, gut microorganisms are important helpers in soil pollution remediation by participating in the preferential degradation of S-IMA mediated by indigenous soil microorganisms.

Comprehensive review on patulin and Alternaria toxins in fruit and derived products.

Mycotoxins are toxic secondary metabolites produced by certain fungi, which can contaminate various food commodities, including fruits and their derived products. Patulin and Alternaria toxins are among the most commonly encountered mycotoxins in fruit and their derived products. In this review, the sources, toxicity, and regulations related to these mycotoxins, as well as their detection and mitigation strategies are widely discussed. Patulin is a mycotoxin produced mainly by the fungal genera Penicillium, Aspergillus, and Byssochlamys. Alternaria toxins, produced by fungi in the Alternaria genus, are another common group of mycotoxins found in fruits and fruit products. The most prevalent Alternaria toxins are alternariol (AOH) and alternariol monomethyl ether (AME). These mycotoxins are of concern due to their potential negative effects on human health. Ingesting fruits contaminated with these mycotoxins can cause acute and chronic health problems. Detection of patulin and Alternaria toxins in fruit and their derived products can be challenging due to their low concentrations and the complexity of the food matrices. Common analytical methods, good agricultural practices, and contamination monitoring of these mycotoxins are important for safe consumption of fruits and derived products. And Future research will continue to explore new methods for detecting and managing these mycotoxins, with the ultimate goal of ensuring the safety and quality of fruits and derived product supply.

Heavy metal contamination and risk assessment in winter jujube (Ziziphus jujuba Mill. cv. Dongzao).

Winter jujube (Ziziphus jujuba Mill. cv. Dongzao) is a major fresh-eating jujube fruit with various important nutrients for humans. It can absorb heavy metals from polluted air, water and soils and applied pesticides, which may pose potential threats to consumers. Here, to evaluate the content of heavy metals in winter jujube and systematically evaluate the potential risks, we collected 212 winter jujube samples from four main producing areas in China and determined the contents of eight heavy metals (Cd, Cr, Pb, Ni, Cu, Zn, As, and Mn) using inductively coupled plasma mass spectrometer (ICP-MS). Based on the integrated pollution index (IPI) evaluation standard, more than 99.06% of samples were at safe levels. Moreover, clustering analysis divided the eight heavy metals into four groups, namely Cr/Ni, Cd/Pb, Cu/Mn/Zn, and As. Importantly, none of the analyzed heavy metals posed risks to adults as indicted by the average carcinogenic and non-carcinogenic risks. Notably, Cr and Cd could pose low carcinogenic risks to children (≤12 age group) when their concentration reached the 90th percentile. This study systematically assessed the health risks associated with heavy metal intake through winter jujube consumption and highlighted the necessity of constant heavy metal monitoring in winter jujube.

Deciphering the diversity, composition, function, and network complexity of the soil microbial community after repeated exposure to a fungicide boscalid.

Boscalid is a novel, highly effective carboximide fungicide that has been substantially and irrationally applied in greenhouses. However, little is known about the residual characteristics of boscalid and its ecological effects in long-term polluted greenhouse soils. Therefore, actual boscalid pollution status in greenhouse soils was simulated by repeatedly introducing boscalid into the soil under laboratory conditions. The degradation characteristics of boscalid, and its effects on the diversity, composition, function, and co-occurrence patterns of the soil microbial community were systematically investigated. Boscalid degraded slowly, with its degradation half-lives ranging from 31.5 days to 180.1 days in the soil. Boscalid degradation was further delayed by repeated treatment and increasing its initial concentration. Boscalid significantly decreased soil microbial diversity, particularly at the recommended dosage. Amplicon sequencing analysis showed that boscalid altered the soil microbial community and further stimulated the phylum Proteobacteria and four potential boscalid-degrading bacterial genera, Sphingomonas, Starkeya, Citrobacter, and Castellaniella. Although the network analysis revealed that boscalid significantly reduced the microbial network complexity, it enhanced the vital roles of Proteobacteria by increasing its proportion and strengthening the relationships among the internal bacteria in the network. The soil microbial function in the boscalid treatment were simulated at the recommended dosage and two-fold recommended dosage but showed an inhibition-recovery-stimulation trend at the five-fold recommended dosage with an increase in treatment frequency. Moreover, the expression of nitrogen cycling functional genes, nifH, AOA amoA, AOB amoA, nirK, and nirS in all boscalid treatments displayed an inhibition-recovery-stimulation trend during the entire experimental period, and the effects were more pronounced at the five-fold recommended dosage. In conclusion, repeated boscalid treatments delayed degradation, reduced soil microbial diversity and network complexity, disturbed soil microbial community, and interfered with soil microbial function.

Systematic evaluation of chiral fungicide penflufen for the bioactivity improvement and input reduction using alphafold2 models and transcriptome sequencing.

Traditional risk assessment of pesticide concludes at the racemic level, which is often incomprehensive. In this study, systematic studies on environmental stability, bioactivity, and ecotoxicological effects of fungicide penflufen were carried out at the enantiomeric level. The single-enantiomer of penflufen was successfully separated and prepared, and their stability was verified in different environmental matrices. Meanwhile, bioactivity test indicated that S-(+)-penflufen had increased bioactivity with its bioactivities against Rhizoctonia solani, Fusarium oxysporum, and Fusarium moniliforme being factors of 7.8, 1.8, and 4.7, respectively greater than those of R-(-)-penflufen. Molecular docking results showed the strong hydrogen bond interactions with Leu300, enantiomer-specific hydrophobic interactions with Cys299, Arg91, and His93, and the greater binding energy between S-(+)-penflufen and succinate dehydrogenase of Rhizoctonia solani caused the selective bioactivity. Additionally, two enantiomers showed low acute toxicity whereas selective sub-chronic toxicity to earthworms. In sub-chronic toxicity test, the accumulated enantiomers caused abnormalities in intestinal tract structure, enzyme activities, and gene expression of earthworms, especially in the S-(+)-penflufen treatment. The selective interactions between penflufen enantiomers and key proteins were elucidated using molecular docking, which may be the main reason of stereoselective subchronic toxicity. S-(+)-penflufen has high bioactivity and low acute risk, it has great potential for development.

Indigenous functional microbial degradation of the chiral fungicide mandipropamid in repeatedly treated soils: Preferential changes in the R-enantiomer.

This study investigated the indigenous functional microbial communities associated with the degradation of chiral fungicide mandipropamid enantiomers in soils repeatedly treated with a single enantiomer. The R-enantiomer degraded faster than the S-enantiomer, with degradation half-lives ranging from 10.2 d to 79.2 d for the R-enantiomer and 10.4 d to 130.5 d for the S-enantiomer. Six bacterial genera, (Burkholderia, Paraburkholderia, Hyphomicrobium, Methylobacterium, Caballeronia, and Ralstonia) with R-enantiomer substrate preference and three bacterial genera (Haliangium, Sorangium, and Sandaracinus) with S-enantiomer substate preference were responsible for the preferential degradation of the R-enantiomer and S-enantiomer, respectively. KEGG analysis indicated that Burkholderia, Paraburkholderia, Hyphomicrobium, and Methylobacterium were the dominant contributors to soil microbial metabolic functions. Notably, six microbial metabolic pathways and twelve functional enzyme genes were associated with the preferential degradation of the R-enantiomer, whose relative abundances in the R-enantiomer treatment were higher than those in the S-enantiomer treatment. A constructed biodegradation gene (BDG) protein database analysis further confirmed that Burkholderia, Paraburkholderia, Hyphomicrobium, Methylobacterium, and Ralstonia were the potential hosts of five dominant BDGs, bphA1, benA, bph, p450, and ppah. We concluded that bacterial genera Burkholderia, Paraburkholderia, Hyphomicrobium, and Methylobacterium may play pivotal roles in the preferential degradation of mandipropamid R-enantiomer in repeatedly treated soils.

Indigenous functional microbial communities for the preferential degradation of chloroacetamide herbicide S-enantiomers in soil.

This study investigated indigenous functional microbial communities associated with the degradation of chloroacetamide herbicides acetochlor (ACE), S-metolachlor (S-MET) and their enantiomers in repeatedly treated soils. The results showed that biodegradation was the main process for the degradation of ACE, S-MET and their enantiomers. Eight dominant bacterial genera associated with the degradation were found: Amycolatopsis, Saccharomonospora, Mycoplasma, Myroides, Mycobacterium, Burkholderia, Afipia, and Kribbella. The S-enantiomers of ACE and S-MET were preferentially degraded, which mainly relied on Amycolatopsis, Saccharomonospora and Kribbella for the ACE S-enantiomer and Amycolatopsis and Saccharomonospora for the S-MET S-enantiomer. Importantly, the relative abundances of Amycolatopsis and Saccharomonospora increased by 146.3%-4467.2% in the S-enantiomer treatments of ACE and S-MET compared with the control, which were significantly higher than that in the corresponding R-enantiomer treatments (25.3%-4168.2%). Both metagenomic and qPCR analyses demonstrated that four genes, ppah, alkb, benA, and p450, were the dominant biodegradation genes (BDGs) potentially involved in the preferential degradation of the S-enantiomers of ACE and S-MET. Furthermore, network analysis suggested that Amycolatopsis, Saccharomonospora, Mycoplasma, Myroides, and Mycobacterium were the potential hosts of these four BDGs. Our findings indicated that Amycolatopsis and Saccharomonospora might play pivotal roles in the preferential degradation of the S-enantiomers of ACE and S-MET.

Repeated exposure to fungicide tebuconazole alters the degradation characteristics, soil microbial community and functional profiles.

Tebuconazole is a broad-spectrum triazole fungicide that has been extensively applied in agriculture, but its toxicity on soil ecology remains unknown after repeated introduction to soil. This study investigated the degradation of tebuconazole and the changes in soil microbial community composition and functional diversity as well as network complexity in soil repeatedly treated with tebuconazole. Tebuconazole degraded slowly as the degradation half-life initially increased and then decreased during the four repeated treatments. High concentration of tebuconazole treatment significantly delayed the degradation of tebuconazole. The soil microbial functional diversity in tebuconazole-treated soils showed an inhibition-recovery-stimulation trend with increasing treatment frequency, which was related to the increased degradation rates of tebuconazole. Tebuconazole significantly decreased soil microbial biomass and bacterial community diversity, and this decreasing trend became more pronounced with increasing treatment frequency and concentration. Moreover, tebuconazole significantly decreased soil bacterial community network complexity, particularly at high concentration of tebuconazole treatment. Notably, four bacterial genera, Methylobacterium, Burkholderia, Hyphomicrobium, and Dermacoccus, were identified as the potential tebuconazole-degrading bacteria, with the relative abundances in the tebuconazole treatment significantly increasing by 42.1-34687.1% compared to the control. High concentration of tebuconazole treatment delayed increases in the relative abundances of Methylobacterium but promoted those of Burkholderia, Hyphomicrobium and Dermacoccus. Additionally, repeated tebuconazole treatments improved only four metabolic pathways, cell motility, membrane transport, environmental information processing, and xenobiotics biodegradation and metabolism, which were associated with the degradation of tebuconazole. The above results indicated that repeated tebuconazole treatments resulted in the significant accumulation of residues and long-term negative effects on soil ecology, and also emphasized the potential roles of dominant indigenous microbial bacteria in the degradation of tebuconazole.

Earthworms accelerated the degradation of the highly toxic acetochlor S-enantiomer by stimulating soil microbiota in repeatedly treated soils.

This study investigated the effects of earthworms on the enantioselective degradation of chloroacetamide herbicide acetochlor with soil microorganisms in repeatedly treated soils. The S-enantiomer degraded more slowly and exerted stronger inhibition on soil microbial functions than the R-enantiomer in single soil system. A synergistic effect was observed between soil microorganisms and earthworms that accelerated the degradation of both the enantiomers, particularly the highly toxic S-enantiomer, which resulted in the preferential degradation of S-enantiomer in soil-earthworm system. Earthworms stimulated five potential indigenous degraders (i.e. Lysobacter, Kaistobacter, Flavobacterium, Arenimonas, and Aquicell), induced two new potential degraders (i.e. Aeromonas and Algoriphagus), and also significantly strengthened the correlations among these seven dominant potential degraders and other microorganisms. Notably, the relative abundances of Flavobacterium and Aeromonas in soil treated with earthworms for S-enantiomer were higher than those for R-enantiomer. Furthermore, earthworms significantly stimulated overall soil microbial activity and improved three microbial metabolic pathways, and xenobiotics biodegradation and metabolism, signal transduction, cell motility, particularly for the S-enantiomer treatment with earthworms, which alleviated the strong inhibition of S-enantiomer on microbial community functions. This study confirmed that earthworms accelerated the degradation of the highly toxic acetochlor S-enantiomer in soil, providing a potential approach in chloroacetamide herbicide-polluted soil remediation.

Enantioselective bioaccumulation and detoxification mechanisms of earthworms (Eisenia fetida) exposed to mandipropamid.

As a novel chiral amide fungicide, the enantioselective behaviors of mandipropamid in the soil environment are unclear. Furthermore, there is a need to understand the stress response mechanisms of soil organisms exposed to mandipropamid isomers. Therefore, the selective bioaccumulation of mandipropamid isomers and detoxification mechanisms of earthworms (Eisenia fetida) were investigated in this study. Our results suggested that the enantioselective bioaccumulation of mandipropamid in earthworms occurred with the preferential enrichment of S-(+)-isomer. The activities of detoxification enzymes, such as cytochrome P450 (CYP450), glutathione-S-transferases (GST), and carboxylesterase (CarE), changed significantly upon exposure to S-(+)- and R-(-)-mandipropamid (particularly for CYP450 and GST). A transcriptome analysis revealed that more differentially expressed genes (DEGs) were observed under S-(+)-isomer exposure (15,798) than those under R-(-)-isomer exposure (12,222), as compared to the control group. These DEGs were mainly enriched in bile secretion and thyroid hormone signaling pathways, which were related to the detoxification process in earthworms. Moreover, the 20 DEGs, which exhibited the most profound changes (such as CYP2 and CYP3A4) in these pathways, were screened, clustered, and observed to be mainly involved in regulating the detoxification function of earthworm cells. These results indicated that detoxification systems played an essential role in the stress response to mandipropamid exposure. Additionally, earthworms were more sensitive to the stress induced by S-(+)-mandipropamid than that induced by R-(-)-mandipropamid. This is the first study to elucidate the mandipropamid detoxification mechanism of earthworms at the enantiomer level, which can be beneficial for remediating chiral pollutants.

Enantioselective residues and toxicity effects of the chiral triazole fungicide hexaconazole in earthworms (Eisenia fetida).

The enantioselective toxic effect and environmental behavior of chiral pesticides have attracted increasing research attention. In this study, the enantioselective toxicity and residues of hexaconazole (HEX) in earthworms (Eisenia fetida) were investigated. In the present study, significant enantioselective degradation characteristics were observed in artificial soil with the R-enantiomer preferentially degrading (p < 0.05); however, no significant enantioselective bioaccumulation was observed in the earthworms (p > 0.05). The acute toxicity of S-(+)-HEX was higher than that of R-(-)-HEX in earthworms, with 48-h LC50 values of 8.62 and 22.35 µg/cm2, respectively. At 25 mg/kg, enantiospecific induction of oxidative stress was observed in earthworms; moreover, S-(+)-HEX had a greater influence on the contents of malonaldehyde, cytochrome P450, and 8-hydroxy-2-deoxyguanosine than R-(-)-HEX. These results were consistent with those of the enrichment analysis of differentially expressed genes. The transcriptome sequencing results showed that S-(+)-HEX had a more significant influence on steroid biosynthesis, arachidonic acid metabolism, and cell cycle processes than R-(-)-HEX, leading to abnormal biological function activities. These results indicate that S-(+)-HEX may pose a higher risk to soil organisms than R-(-)-HEX. This study suggests that the environmental risk of chiral pesticides to nontarget organisms should be assessed at the enantiomeric level.

Dissipation of chlorothalonil in the presence of chlortetracycline and ciprofloxacin and their combined effects on soil enzyme activity.

The long-term application of substantial amounts of fungicides and antibiotic-polluted organic manure (OM) in greenhouse has caused the co-existence of fungicides and antibiotics in soils. However, little is known about the effects of antibiotics on the persistence of fungicides in soils or their combined effects on soil enzyme activity. In this study, fungicide chlorothalonil (CTL) alone and in combination with antibiotic chlortetracycline (CTC) or ciprofloxacin (CIP) were repeatedly added to OM-amended soil to investigate the changes in the residual characteristics of CTL and in soil dehydrogenase and urease activity. The results showed that CTL rapidly dissipated in soils with the corresponding half-lives of 0.9-3.2, which initially increased, then decreased and finally stabilized with an increased treatment frequency. The dissipation of CTL was inhibited by CTC and CIP during the first several treatments. The soil dehydrogenase and urease activity in CTL-treated soils was inhibited during the first six treatments and then recovered afterwards. Compared with the OM-amended soil+CTL treatment, the OM-amended soil+CTL+CTC and OM-amended soil+CTL+CIP treatments had stronger inhibitory effects on soil enzyme activity during the first six repeated treatments but exhibited slight stimulating effects afterwards. Therefore, the results obtained in this study suggested that the long-term co-existence of CTL, CTC, and CIP altered the dissipation characteristics of CTL in soil and affected the soil enzyme activity levels. The prudent application of large and frequent of fungicides and OM-containing antibiotic residues in greenhouses should therefore be carefully considered in order to reduce the long-term combined pollution in soils.

Azoxystrobin dissipation and its effect on soil microbial community structure and function in the presence of chlorothalonil, chlortetracycline and ciprofloxacin.

The residual characteristics and the adsorption-desorption behaviors of azoxystrobin (AZO) as well as the soil ecological effects in the individual repeated treatments of AZO and its combination with chlorothalonil (CTL), chlortetracycline (CTC) and ciprofloxacin (CIP) were systematically studied in organic manure (OM)-amended soil under laboratory conditions. The presence of CTL, CTC, and CIP, both individually and combined, decreased the sorption affinity of AZO with the Freundlich adsorption and desorption coefficient decreasing by 0.3-24.2%, and CTC and CIP exhibited greater adverse effects than CTL. AZO dissipated slowly and the residues significantly accumulated during ten repeated treatments. The dissipation of AZO was inhibited to different degrees in the combined treatments. Biolog analysis revealed that the soil microbial functional diversity in the OM-soil + AZO and OM-soil + AZO + CTL treatments was higher than that in the OM-soil treatment during the former three repeated treatments, but which was inhibited during the latter seven repeated treatments. The soil microbial functional diversity in the OM-soil + AZO + CTC, OM-soil + AZO + CIP and OM-soil + AZO + CTL + CTC + CIP treatments was inhibited during the ten repeated treatments compared with OM-soil treatment. Metagenomic results showed that all repeated treatments significantly increased the relative abundance of Actinobacteria, but significantly decreased that of Proteobacteria and Firmicutes during the ten repeated treatments. Furthermore, the relative abundance of soil dominant bacterial genera Rhodococcus, Mycobacterium and Arthrobacter in all the repeated treatments significantly increased by 1.5-1283.9% compared with the OM-soil treatment. It is concluded that coexistence of CTL, CTC and CIP, both individually and combined, with AZO can inhibit the dissipation of AZO, reduce the adsorption affinity of AZO on soil, and alter the soil microbial community structure and functional diversity.

Deposition, Distribution, Metabolism, and Reduced Application Dose of Thiamethoxam in a Pepper-Planted Ecosystem.

To decrease the application dose of thiamethoxam (TMX) to control the pepper whitefly (Bemisia tabaci Q), the deposition, dissipation, metabolism, and field efficacy of TMX were investigated in a pepper (Capsicum annuum var. grossum)-planted ecosystem using eight types of nozzles at six concentrations (56.25, 41.25, 26.25, 21.0, 15.75, and 10.5 g a.i./hm2). The initial deposition amount of TMX in the pepper plant first increased and then decreased with increasing application dose. The optimum spray conditions of TMX were found to be a droplet size of 200 µm volume median diameter and a spray volume of 350 L/hm2. Moreover, three metabolites, TMX-dm, clothianidin (CLO), and C5H8O2N3SCl, were detected in the pepper-planted system. The dissipation rate of TMX in the pepper-field ecosystem was leaves > stems > fruits > roots > soils. The results revealed the deposition and fate of TMX in the pepper-field ecosystem, and the application dose could be reduced by 20% based on the minimum recommended dose for controlling pepper whitefly.

Development of antibiotic resistance genes in soils with ten successive treatments of chlortetracycline and ciprofloxacin.

Antibiotic contamination caused by the long-term use of organic manure (OM) in greenhouse agricultural soils poses potential detrimental effects to the soil environment. By applying OM containing chlortetracycline (CTC) and/or ciprofloxacin (CIP) ten times in soil under laboratory conditions, we investigated the dissipation and accumulation characteristics of CTC and CIP in the soil, the changes in the microbial pollution-induced community tolerance (PICT), and the diversity and abundance of antibiotic resistance genes (ARGs) in the soil microbiome. The dissipation of CTC was rapid while CIP was accumulated in repeatedly treated soils; further, CIP could inhibit the dissipation of CTC. Meanwhile, the PICT to CTC and/or CIP significantly increased up to 15.0-fold after ten successive treatments compared to that in the first treatment. As the treatment frequency increased, significant upward trends in the abundances of tetracycline resistance genes tetA(G), tetX2, tetX, tetG, tetA(33), tetA, tetW, and tetA(P), fluoroquinolone resistance gene qnrA6, and multiple resistance gene mexF were revealed by both metagenomic and qPCR analyses. The findings demonstrated that repeated treatments with CTC and/or CIP can alter the dissipation rate, promote an increase in PICT to CTC and/or CIP, and increase the ARGs abundance in steps.

Exploring microbial community structure and biological function in manured soil during ten repeated treatments with chlortetracycline and ciprofloxacin.

The changes of enzyme activities, microbial community structure and function, and the diversity and resistance level of antibiotic-resistant bacteria (ARB) were studied in soil during ten repeated treatments with chlortetracycline (CTC) and/or ciprofloxacin (CIP) together with organic manure (OM) under laboratory conditions. The activities of neutral phosphatase (NPA) and catalase (CAT) displayed the suppression-recovery-stimulation trend in the OM&CTC treatment but the stimulation trend in the OM&CTC&CIP treatment. The NPA was stimulated but the CAT was little affected in the OM&CIP treatment. Soil microbial functional diversity displayed the suppression-recovery-stimulation trend in the OM&CTC and OM&CTC&CIP treatments and the stimulation-suppression trend in the OM&CIP treatment with the treatment frequency. Metagenomic analysis showed that the relative abundances of Actinobacteria and Firmicutes in the antibiotic treatment significantly increased by 0.5-235.6%, but that of Proteobacteria decreased by 0.2-27.3% compared to the control with the treatment frequency. Furthermore, the relative abundances of dominant bacterial genera including Streptomyces, Actinomadura, Mycobacterium, and Streptococcus in the antibiotic treatment significantly increased by 1.1-10433.3% compared to the control. Meanwhile, repeated antibiotic treatments induced a significant increase in the diversity and resistance level of ARB isolates, especially in the OM&CTC treatment. It is concluded that repeated treatments with CTC and/or CIP can alter enzyme activities, microbial community structure and function, and increase the diversity and resistance level of ARB isolates.