Epoxiconazole disturbed metabolic balance and gut microbiota homeostasis in juvenile zebrafish.

Epoxiconazole (EPX) is a broad-spectrum fungicide extensively used in agricultural pest control. Emerging evidence suggests that EPX can adversely affect different endpoints in non-target organisms. Here, the toxicity of EPX was assessed using earlier developmental stage of zebrafish as a model to investigate its effects on metabolism and intestinal microbiota after 21 days of exposure. Our findings indicated that EPX exposure resulted in physiological alterations in juvenile zebrafish, including increase in triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL), and glycose (Glu). Nile red staining demonstrated enhanced lipid accumulation in juvenile, accompanied by a marked upregulation in the expression of genes associated with TG synthesis. Moreover, EPX led to alterations in amino acids and carnitines levels in 21 dpf (days post fertilization) zebrafish. We also observed that EPX disrupted intestinal barrier function in juvenile zebrafish, manifested by decreasing mucus secretion and changing in genes related to tight junctions. Moreover, for a more comprehensive analysis of the intestinal microbiota in 21 dpf zebrafish, the intestine tissues were dissected under a microscope for 16S rRNA sequencing analysis. The results revealed that EPX altered the structure and abundance of intestinal microflora in zebrafish, including decreased alpha diversity indices and shifted in bacteria at phylum and genus levels. Notably, the correlation analysis demonstrated strong associations between alterations in various pathogenic bacterial genera and levels of amino acids and carnitines. Overall, these findings confirm that the fungicide EPX promotes metabolic disorders and alterations in the intestinal micro-environment in 21 dpf zebrafish, shedding light on the toxicologic effects of chemicals to aquatic organisms during the development stage.

Epoxiconazole altered hepatic metabolism in adult zebrafish based on transcriptomic analysis.

Epoxiconazole (EPX) is a triazole fungicide, which has been widely used in pest control of cereal crops. However, its extensive use has led to concerning levels of residue in water bodies, posing substantial risks to aquatic life. In this study, we characterized the toxicological effects of EPX on 6-month-old male and female zebrafish at 70 and 700 µg/L, respectively. The results revealed that EPX exposure markedly increased both body length and weight in zebrafish of both sexes, consequently elevating their condition factor. Besides, EPX exposure resulted in notable alterations in hepatic histopathology. These changes included loosened hepatocyte structure, ballooning degeneration, nucleolysis, and disappearance of cell line, with male zebrafish exhibiting more severe damage. High concentration of EPX also significantly increased hepatic lipid accumulation in male zebrafish, as well as increased hepatic triglyceride (TG) levels. Correspondingly, there was a notable alteration in the transcription of genes including cyp51, hmgcr, and PPAR-γ, which associated with cholesterol and lipid metabolism. Interestingly, with the hepatic transcriptomic analysis, high concentration of EPX produced 195 upregulated and 107 downregulated differential expression genes. Both KEGG and GO analyses identified significant enrichment of these genes in lipid and amino acid metabolism pathways. Notably, some key genes involved in the steroid synthesis pathway were marked upregulated. In addition, molecular docking study confirmed that EPX could bind CYP51 protein well (△G = -7.7 kcal/mol). Taken together, these findings demonstrated the multiple toxic effects of EPX on adult zebrafish.

Toxicologic effect and transcriptome analysis for sub-chronic exposure to carbendazim, prochloraz, and their combination on the liver of mice.

Carbendazim (CBZ) and prochloraz (PCZ) are broad-spectrum fungicides used in agricultural peat control. Both fungicides leave large amounts of residues in fruits and are toxic to non-target organisms. However, the combined toxicity of the fungicides to non-target organisms is still unknown. Therefore, we characterized the toxic effects of dietary supplementation with CBZ, PCZ, and their combination for 90 days in 6-week-old male Institute of Cancer Research (ICR) mice. CBZ-H (100 mg/kg day), PCZ-H (10 mg/kg day), and their combination treatments increased the relative liver weights and caused liver injury. The serum total cholesterol (TC), triglyceride (TG), glucose (Glu), pyruvate (PYR), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels were reduced, and synergistic toxicity was observed. Hepatic transcriptome revealed that 326 differentially expressed genes (DEGs) of liver were observed in the CBZ treatment group, 149 DEGs in the PCZ treatment group, and 272 DEGs in the combination treatment group. According to KEGG enrichment analysis, the fungicides and their combination affected lipid metabolism, amino acid metabolism, and ferroptosis. In addition, the relative mRNA levels of key genes involved in lipid metabolism were also examined. Compared with individual exposure, combined exposure to CBZ and PCZ caused a more obvious decrease in the expression of some genes related to glycolipid metabolism. Furthermore, the relative mRNA levels of some key genes in the combination treatment group were lower than those in the CBZ and PCZ treated groups. In summary, CBZ, PCZ, and their combination generally caused hepatotoxicity and glycolipid metabolism disorders, which could provide new insights for investigating the combined toxicity of multiple fungicides to animals.

Oral Exposure to Epoxiconazole Disturbed the Gut Micro-Environment and Metabolic Profiling in Male Mice.

Epoxiconazole (EPX), a triazole fungicide, is widely used in agriculture to control pests and diseases. High residual and occupational exposure to EPX increases health risks, and evidence of potential harm to mammals remains to be added. In the present study, 6-week-old male mice were exposed to 10 and 50 mg/kg bw EPX for 28 days. The results showed that EPX significantly increased the liver weights. EPX also decreased the mucus secretion of the colon and altered intestinal barrier function in mice including a reduced expression of some genes (Muc2, meprinß, tjp1). Moreover, EPX altered the composition and abundance of gut microbiota in the colon of mice. The alpha diversity indices (Shannon, Simpson) in the gut microbiota increased after exposure to EPX for 28 days. Interestingly, EPX increased the ratio of Firmicutes to Bacteroides and the abundance of other harmful bacteria including Helicobacter and Alistipes. Based on the untargeted metabolomic analysis, it was found that EPX altered the metabolic profiles of the liver in mice. KEGG analysis of differential metabolites revealed that EPX disrupted the pathway related to glycolipid metabolism, and the mRNA levels of related genes were also confirmed. In addition, the correlation analysis showed that the most altered harmful bacteria were associated with some significantly altered metabolites. The findings highlight that EPX exposure changed the micro-environment and lipid metabolism disturbance. These results also suggest that the potential toxicity of triazole fungicides to mammals cannot be ignored.

Antimicrobial and the Resistances in the Environment: Ecological and Health Risks, Influencing Factors, and Mitigation Strategies.

Antimicrobial contamination and antimicrobial resistance have become global environmental and health problems. A large number of antimicrobials are used in medical and animal husbandry, leading to the continuous release of residual antimicrobials into the environment. It not only causes ecological harm, but also promotes the occurrence and spread of antimicrobial resistance. The role of environmental factors in antimicrobial contamination and the spread of antimicrobial resistance is often overlooked. There are a large number of antimicrobial-resistant bacteria and antimicrobial resistance genes in human beings, which increases the likelihood that pathogenic bacteria acquire resistance, and also adds opportunities for human contact with antimicrobial-resistant pathogens. In this paper, we review the fate of antimicrobials and antimicrobial resistance in the environment, including the occurrence, spread, and impact on ecological and human health. More importantly, this review emphasizes a number of environmental factors that can exacerbate antimicrobial contamination and the spread of antimicrobial resistance. In the future, the timely removal of antimicrobials and antimicrobial resistance genes in the environment will be more effective in alleviating antimicrobial contamination and antimicrobial resistance.

Maternal procymidone exposure has lasting effects on murine gut-liver axis and glucolipid metabolism in offspring.

There is increasing evidence that maternal exposure to environmental pollutants can cause intestinal and metabolic diseases, and these disease risks still exist in offspring. Here, female C57BL/6 mice were orally treated with procymidone (PRO) (10 and 100 mg/kg body weight/day) by dietary supplementation during the gestation and lactation periods. Then, we discovered PRO changed the physiology, intestinal barrier and metabolism both in the generations of F0 and different developmental stages of F1 (7 weeks and 30 weeks old, respectively). Maternal PRO exposure affected the growth phenotypes and the glucolipid metabolism related indicators and genes of mice, especially the male mice of F1 generations. The changes in bile acids (BAs) metabolism demonstrated that PRO disordered glucolipid metabolism through enterohepatic circulation. Furthermore, PRO reduced mucus secretion in the gut and altered the composition of gut microbiota, leading more bacteria to disseminate in the gut and inflammatory responses both in F0 and F1 regenerations. And PRO-induced gut microbiota dysbiosis was tightly related to BAs metabolites. Together, the results indicated that PRO destructed the functional integrity of intestinal barrier and the inflammatory reaction was triggered. And then, the disorder of glucolipid metabolism was induced through the BAs enterohepatic circulation. This study indicated that the cross-generation effects of PRO could not be ignored.

Effect of High-Pressure GaN Nucleation Layer on the Performance of AlGaN/GaN HEMTs on Si Substrate.

A high-pressure (HP) GaN nucleation layer (NL) was inserted between AlGaN buffer and an unintentionally doped (UID) GaN layer of an AlGaN/GaN HEMT on Si. The XRD and TEM showed that when the V/III ratio was optimized during the HP-GaN NL growth, the edge dislocation density in the HP-GaN NL layer could be reduced significantly. Experimental results exhibited a lower off-state leakage current, higher maximum ID and Gm (corresponding to 22.5% and 21.7% improvement, respectively), and lower on-state resistance. These results demonstrate that the electrical properties of the AlGaN/GaN HEMT can be improved through the insertion of a HP-GaN NL.

Nano- and micro-polystyrene plastics interfered the gut barrier function mediated by exosomal miRNAs in rats.

Microplastics (MPs) are widely distributed in the global environment, entering and accumulating in organisms in various ways and posing health threats. MPs can damage intestine; however, the mechanism by which MPs cause intestinal damage in rats is unclear. Here, rats were exposed to 50 nm PS-NPs or 5 µm PS-MPs for 4 weeks to evaluate the possible effects on intestinal barrier function and exosomal miRNAs expressions. The results showed that PS-NPs or PS-MPs disrupted the gut microbiota and affected gut barrier function at the biological level. In addition, PS-NPs and PS-MPs altered the composition of exosomal miRNAs in the intestinal and serum. Both PS-NPs and PS-MPs decreased the expression of miR-126a-3p in the intestinal and serum exosomes, which is an important signalling molecule involved in MPs induced gut barrier function disorder. More importantly, both in vitro and in vivo experiments indicated that miR-126a-3p was closely related to oxidative damage of intestinal cells through the PI3K-Akt pathway and eventually promote cell apoptosis by regulating the target gene of PIK3R2. Our study suggested that PS-NPs and PS-MPs could affect rat intestinal barrier function through an exosomal miRNA mediated pathway.

Impact of Radiotherapy Combined With Chemotherapy on Long-Term Outcomes of Patients With Recurrent Nasopharyngeal Carcinoma.

Background: It remains controversial whether the application of chemotherapy has an impact on recurrent nasopharyngeal carcinoma (rNPC) patients with salvage radiotherapy. Here, we aimed to evaluate treatment outcomes of rNPC patients and derive a prognostic model to assess the benefit of chemotherapy in patients with re-radiotherapy. Methods: This study was conducted as a retrospective study. In total, 340 rNPC patients treated with salvage intensity-modulated radiotherapy (IMRT) or radiochemotherapy (RCT) from October 2006 to September 2019 were included in this study. Overall survival (OS) was the primary outcome. Kaplan-Meier method was employed to detect the prognostic difference with Log-rank tests. The Cox regression analysis was performed to explore the potential prognostic factors while the multivariate Cox analysis was used to identify candidate variables for the prognostic model of OS. Results: The 5-year actuarial rates of OS, progression-free survival, loco-regional progression-free survival, and distant metastases-free survival did not show significant difference between the IMRT and RCT groups (P > .05). Age at recurrence and rT category were found to be the independent prognostic factors for OS. We found that rNPC patients suffered poor OS in the high-risk group (patients with higher age at recurrence and advanced rT category) (high-risk vs low-risk, HR = 1.87, 95% CI: 1.36-2.57, P < .001). Salvage RT alone may be superior to RCT for patients in the low-risk group (RCT group vs RT group, HR = 1.89, 95% CI: 1.11-3.20, P = .038). Conclusion: Salvage RT combined with chemotherapy cannot improve survival outcomes for rNPC. More novel clinical trials should be explored to develop individualized strategies to improve survival and minimize toxicities.

Transcriptomic and targeted metabolomic analysis revealed the toxic effects of prochloraz on larval zebrafish.

Prochloraz (PCZ), an imidazole fungicide, has been extensively used in horticulture and agriculture to protect against pests and diseases. To investigate the potential toxicity of PCZ on aquatic organisms, larval zebrafish, as a model, were exposed to a series of concentrations (0, 20, 100, and 500 µg/L) of PCZ for 7 days. With transcriptomic analysis, we found that exposure to high dose PCZ could produce 76 downregulated and 345 upregulated differential expression genes (DEGs). Bioinformatics analysis revealed that most of the DEGs were characterized in the pathways of glycolipid metabolism, amino acid metabolism and oxidative stress in larval zebrafish. Targeted metabolomic analysis was conducted to verify the effects of PCZ on the levels of acyl-carnitines and some amino acids in larval zebrafish. In addition, biochemical indicators related to glycolipid metabolism were affected obviously, manifested as elevated triglyceride (TG) levels and decreased glucose (Glu) levels in whole larvae. The expression levels of genes associated with glycolipid metabolism were affected in larvae after exposure to PCZ (PK, GK, PEPckc, SREBP, ACO). Interestingly, we further confirmed that PCZ could induce oxidative stress by the changing enzyme activities (T-GSH, GSSG) and upregulating several related genes levels in larval zebrafish. Generally, our results revealed that the endpoints related to glycolipid metabolism, amino acid metabolism and oxidative stress were influenced by PCZ in larval zebrafish.

Bromuconazole exposure induces cardiotoxicity and lipid transport disorder in larval zebrafish.

Bromuconazole (BRO), as one of the typical triazole fungicides, has not been reported on its effects on aquatic organisms. In this study, zebrafish embryos were used as experimental objects to evaluate the toxicity of BRO. In the acute embryo toxicity test, it was observed that the heart rate and growing development were affected by BRO in a concentration-dependent manner, and the half-lethal concentration (LC50) of BRO at 96 h post-fertilization (hpf) was about 11.83 mg/L. Then, low concentrations of BRO (50 ng/L, 0.075 mg/L, 0.3 mg/L, 1.2 mg/L), which were set according to the LC50 and environmental related concentrations, were used to analyze the toxic effects on the different endpoints in larval zebrafish. Interestingly, the transcriptomic analysis found that most different expressed genes (DEGs) could be focused on the pathways of lipid metabolism, myocardial function, glycometabolism, indicating that heart function and lipid metabolism in larval zebrafish were disrupted by BRO. For supporting this idea, we re-exposed the transgenic zebrafish and WT zebrafish embryos, proved that BRO caused damage to heart development and lipid transport on morphological and genetic level, which was consistent with transcriptomic results. In addition, BRO exposure caused oxidative damage in the larvae. Taken together, BRO exposure could affect the myocardial contraction function and lipid transport in larval zebrafish, accompanied by disturbances in the level of oxidative stress, which was of great significance for improving the biotoxicological information of BRO.

Transgenerational effects of co-exposure to cadmium and carbofuran on zebrafish based on biochemical and transcriptomic analyses.

The combined toxicity of heavy metals and pesticides to aquatic organisms is still largely unexplored. In this study, we investigated the combined impacts of cadmium (Cd) and carbofuran (CAR) on female zebrafish (F0 generation) and their following F1 generation. Results showed that mixtures of Cd and CAR induced acute synergistic effects on both zebrafish adults of the F0 generation and embryos of the F1 generation. Combined exposure to Cd and CAR could obviously alter the hepatic VTG level of females, and the individual exposures increased the relative mRNA levels of vtg1 and vtg2. Through maternal transmission, co-exposure of Cd and CAR caused toxicity to 4-day-old larvae of the F1 generation, evidenced by the significant changes in T4 and VTG levels, CYP450 activity, and the relative transcriptional levels of genes related to the hormone, oxidative stress, and apoptosis. These effects were also reflected by the global gene expression pattern to 7-day-old larvae of F1 generation using the transcriptomic analysis, and they could also affect energy metabolism. Our results provided a more comprehensive insight into the transgenerational toxic impacts of heavy metal and pesticide mixtures. These findings highlighted that it was highly necessary to consider transgenerational exposures in the ecological risk assessment of chemical mixtures.

Insights into the effects of difenoconazole on the livers in male mice at the biochemical and transcriptomic levels.

Difenoconazole (DFZ) is a broad-spectrum triazole fungicide, that is extensively used in agriculture. Studies have shown that residues of DFZ and other fungicides have toxic effects on nontarget organisms. However, its hepatoxicity in mammals remains unclear. Here, we characterized the toxic hepatic effects in male C57BL/6 mice exposed to 30 and 100 mg/kg bw DFZ for 14 and 56 days, respectively. The results revealed that DFZ could increase the relative liver weights, however, the relative fat and spleen weights decreased. More importantly, DFZ exposure changed the hepatic morphology and induced hepatic oxidative stress. Gene expression analysis suggested that DFZ could induce a glycolipid metabolism disorder. Moreover, hepatic transcriptomic analysis revealed the effects of DFZ exposure on the transcriptional levels of various genes, and enrichment analysis of differentially expressed genes (DEGs) showed that energy metabolism and immune-associated pathways were mainly affected. We validated the results from transcriptomic analysis and found that some key genes related to energy metabolism were affected. In addition, flow cytometry showed that the CD3+/CD4+ and CD3+ /CD8+ levels declined in the spleen of mice. Taken together, these findings combined with transcriptome analysis highlighted that DFZ caused different endpoints in the liver, which could provide more evidence for investigating the toxic effects of DFZ in mammals.

Buffer Traps Effect on GaN-on-Si High-Electron-Mobility Transistor at Different Substrate Voltages.

Substrate voltage (VSUB) effects on GaN-on-Si high electron mobility transistors (HEMTs) power application performance with superlattice transition layer structure was investigated. The 2DEG conductivity and buffer stack charge redistribution can be affected by neutral/ionized donor and acceptor traps. As the donor/acceptor traps are excessively ionized or de-ionized by applying VSUB, the depletion region between the unintentionally doped (UID)/Carbon-doped (C-doped) GaN layer may exhibit a behavior similar to the p-n junction. An applied negative VSUB increases the concentration of both the ionized donor and acceptor traps, which increases the breakdown voltage (BV) by alleviating the non-uniform distribution of the vertical electric field. On the other hand, an applied positive VSUB causes the energy band bending flattener to refill the ionized traps and slightly improves the dynamic Ron degradation. Moreover, the amount of electrons injected into the buffer stack layer from the front side (2DEG channel/Ohmic contact) and the back side (AlN nucleation layer/superlattice transition layer) are asymmetric. Therefore, different VSUB can affect the conductivity of 2DEG through the field effect, buffer trapping effect, and charge redistribution, which can change the electrical performance of the device.

A Novel GaN:C Millimeter-Wave HEMT with AlGaN Electron-Blocking Layer.

An AlGaN/GaN/Si high electron mobility transistor (HEMT) using a GaN:C buffer with a 2 nm AlGaN electron-blocking layer (EBL) is investigated for the first time for millimeter-wave applications. Compared with the double heterostructure field effect transistor (DHFET), the AlGaN/GaN HEMT with the GaN:C/EBL buffer has a lower vertical leakage, higher thermal stability, and better RF performance. In addition, AlGaN EBL can prevent carbon-related traps from GaN:C and improve electron confinement in 2DEG during high-frequency operation. Finally, a Pout of 31.2 dBm with PAE of 21.7% were measured at 28 GHz at 28 V. These results demonstrated the great potential of HEMTs using GaN:C with AlGaN EBL epitaxy technology for millimeter-wave applications.

Combined hepatotoxicity of imidacloprid and microplastics in adult zebrafish: Endpoints at gene transcription.

Microplastics (MPs) and pesticides are two kinds of ubiquitous pollutants that can pose a health risk to aquatic organisms. However, researches about the combined effects of MPs and pesticides are very limited. A simple combined exposure model was established in this study, adult zebrafish were exposed to 100 µg/L imidacloprid (IMI), 20 µg/L polystyrene microplastics (PS), and a combination of PS and IMI (PS + IMI) for 21 days. The results demonstrated that exposure to PS and IMI inhibited the growth of zebrafish and altered the levels of glycolipid metabolism and oxidative stress-related biochemical parameters. While gene expression analysis revealed that, compared with PS or IMI treatment group, combined exposure caused a greater change in gene expression levels involving the process of glycolipid metabolism (Gk, Hk1, Aco, PPar-α, Cpt1, Acc, Fas, PPar-γ, Apo) and inflammatory response (IL-1ß, IL-6, IL-8, TNF-α, IL-10). The results demonstrated that even combined exposure of low concentrations of PS and IMI could cause more severe hepatotoxicity in zebrafish, especially in terms of gene transcription. And more combined toxicity studies are essential for MPs and pesticides risk assessment.

Maternal exposure to sodium ρ-perfluorous nonenoxybenzene sulfonate during pregnancy and lactation disrupts intestinal barrier and may cause obstacles to the nutrient transport and metabolism in F0 and F1 generations of mice.

Sodium ρ-perfluorous nonenoxybenzene sulfonate (OBS), a novel kind of perfluoroalkyl and polyfluoroalkyl compound, has been widely detected in the environment. The toxicity of OBS to living organisms has become a public concern. A growing body of research showed that maternal exposure to environmental pollutants caused intestinal and metabolic diseases that could be conserved across offspring. Here, female C57BL/6 mice were treated OBS at dietary levels of 0.0 mg/L (CON), 0.5 mg/L (OBS-L) and 5.0 mg/L (OBS-H) during the gestation and lactation periods. The results demonstrated that OBS treatment not only induced significant changes in the mucus secretion and ionic transport, but also disrupted the expression of antimicrobial peptides (AMPs) in the intestine of F0 and F1 generations. Additionally, OBS exposure altered bile acids metabolism and affected the transcriptional levels of critical genes involved in bile acids synthesis, signaling transfer, transportation and apical uptake. Together, all these results indicated that OBS exposure was perceived as a major stress by the intestinal epithelium that strongly affected the intestinal barrier function (including mucus, CFTR, AMPs, inflammation), and ultimately led to imbalance in the metabolism of bile acids (BAs). Moreover, we found that maternal OBS exposure had a more obvious toxicity effect on the male offspring in this experiment. Taken together, maternal OBS exposure during pregnancy and lactation had the intestinal and metabolism toxic effects on the dams and offspring, indicating that effects of maternal exposure on the toxicity of offspring could not be ignored.

Microplastic: A potential threat to human and animal health by interfering with the intestinal barrier function and changing the intestinal microenvironment.

Plastics are widely used in many fields due to their stable physical and chemical properties, and their global production and usage increase significantly every year, which leads to the accumulation of microplastics in the entire ecosystem. Numerous studies have shown that microplastics (MPs) have harmful effects on living organisms. This review aims to provide a comprehensive conclusion of the current knowledge of the impacts of MPs on the stability of the gut microenvironment, especially on the gut barrier. Studies showed that exposure to MPs could cause oxidative damage and inflammation in the gut, as well as the destruction of the gut epithelium, reduction of the mucus layer, microbial disorders, and immune cell toxicity. Although there are few reports directly related to humans, we hoped that this review could bring together more and more evidence that exposure to MPs results in disturbances of the intestinal microenvironment. Therefore, it is necessary to investigate their threats to human health further.

Propamocarb exposure has the potential to accelerate the formation of atherosclerosis in both WT and ApoE-/- mice accompanied by gut microbiota dysbiosis.

Propamocarb is a systemic carbamate fungicide used to fight diseases. The effect of propamocarb on the formation of atherosclerosis was evaluated in wild-type (WT) and ApoE knockout (ApoE-/-) mice. C57BL/6 J WT mice were fed control diet or high-fat diet (HFD) with 20 mg/L propamocarb in drinking water for 24 weeks. Propamocarb significantly increased the serum levels of triglyceride, cholesterol and low-density lipoprotein cholesterol while decreasing high-density lipoprotein cholesterol. Simultaneously, propamocarb facilitated lipid accumulation in the liver and increased the expression of cholesterol synthesis and transport genes in the liver and ileum. Lipid accumulation was observed in the aortic roots of the propamocarb-treated mice fed HFD, and similar results were also observed with whole aorta staining. In addition, propamocarb exposure significantly increased the mRNA levels of IL-1ß, TNF-α, ICAM-1, and VCAM-1 in the aorta and the serum IL-1ß, IL-6, and TNF-α levels in HFD groups treated with propamocarb. In ApoE-/- mice, the results were consistent with those obtained in WT mice after exposure to 20 mg/L propamocarb for 10 weeks. Meanwhile, propamocarb significantly increased the levels of CD36, NF-κB, VCAM-1 and ICAM-1 proteins in the aortas of ApoE-/- mice. Propamocarb further disrupted cholesterol metabolism and enhanced atherosclerosis and inflammatory responses much more substantially, indicating that propamocarb has the potential to accelerate the formation of atherosclerosis. An analysis of gut microbiota revealed that propamocarb altered the composition of gut microbiota in both WT and ApoE-/- mice. Interestingly, propamocarb increased the abundance of Peptostreptococcaceae, Ruminococcaceae, and Clostridiales_VadinBB60_group, which are related to atherosclerosis at the family level. The abundance of Paeniclostridium, Allobaculum, and Clostridioides, which are closely related to atherosclerosis, was also increased by propamocarb exposure. Our findings indicate that propamocarb exposure may promote atherosclerosis by disrupting lipid metabolism, increasing the inflammatory response, and altering the structure of gut microbiota.

Embryonic toxicity of epoxiconazole exposure to the early life stage of zebrafish.

Epoxiconazole (EPX), as a broad-spectrum triazole fungicide, is widely used in agriculture to resist pests and diseases, while it may have potential toxicity to non-target organisms. In the present study, early developmental stage zebrafish were used as the subject organisms to assess the toxicity of EPX, and the possible mechanism of toxicity was also discussed by biochemical and transcriptomic analysis. Through embryo toxicity test, we had made it clear that the 96 h LC50 of embryo was 7.204 mg/L, and acute exposure to EPX effected hatching rate, heartbeats, body length and even morphological defects. Then, by being exposed to EPX for 7 days at concentrations of 175 (1/40 LC50), 350 (1/20 LC50) and 700 (1/10 LC50), biochemical parameters were affected, mainly manifested as increase of the triglyceride (TG) level and decrease of glucose content. Correspondingly, the transcription of genes related of glucose metabolism, lipid metabolism and cholesterol metabolism were also affected significantly in larval zebrafish. Moreover, some pathways, including lipid metabolism, glucose metabolism and amino acid metabolism were affected through transcriptome sequencing analysis in the larval zebrafish. Further data analysis based on the sequencing, EPX exposure also affected the expression of genes related to cell apoptosis. We further conformed that the bright fluorescence on the liver and bright spots near the liver by acridine orange staining. In addition, the mRNA levels of apoptosis related genes were also significantly affected in the EPX exposed larval zebrafish. Taken together, the work could provide an insight into toxic effects of EPX on the zebrafish larvae at embryo toxicity and transcriptional levels, providing some evidences for the toxic effects of triazole fungicides on non-target organisms.