Insights into the developmental and cardiovascular toxicity of bixafen using zebrafish embryos and larvae.

Bixafen (BIX), a member of the succinate dehydrogenase inhibitor (SDHI) class of fungicides, has seen a surge in interest due to its expanding market presence and positive development outlook. However, there is a growing concern about its potential harm to aquatic life, largely due to its resistance to breaking down in the environment. In this study, we thoroughly examined the toxicological impact of BIX on zebrafish as a model organism. Our results revealed that BIX significantly hindered the development of zebrafish embryos, leading to increased mortality, hatching failures, and oxidative stress. Additionally, we observed cardiovascular abnormalities, including dilated cardiac chambers, reduced heart rate, sluggish blood circulation, and impaired vascular function. Notably, BIX also altered the expression of key genes involved in cardiovascular development, such as myl7, vmhc, nkx2.5, tbx5, and flt1. In summary, BIX was found to induce developmental and cardiovascular toxicity in zebrafish, underscoring the risks associated with SDHI pesticides and emphasizing the need for a reassessment of their impact on human health. These findings are crucial for the responsible use of BIX.

Bixafen causes hepatotoxicity and pancreas toxicity in zebrafish (Danio rerio).

Bixafen (BIX), a widely used succinate dehydrogenase inhibitor (SDHI) in agricultural disease control, has garnered significant attention due to its known hazardous effects on aquatic organisms. In this study, we exposed zebrafish embryos to 0.1, 0.2, and 0.3 µM BIX, to explore the impact of BIX on liver and pancreas. The results showed that BIX caused deformities and dysfunction in zebrafish embryos, including spinal curvature, pericardial edema, heart rate decrease, and hatching delay. Moreover, BIX significantly affected the development of the liver and pancreas in zebrafish and downregulated zebrafish fabp10a gene expression. Overall, this study presents strong evidence for BIX's potential toxicity to zebrafish liver and pancreas. The results may provide new insights into the evaluation of BIX'S impact on aquatic organisms.

Realistic exposure of the fungicide bixafen in soil and its toxicity and risk to natural earthworm populations after multiyear use in cereal.

A comprehensive multiyear monitoring program was conducted to assess the exposure, effects, and long-term risk of the fungicide bixafen to earthworms in cereal fields. The realistic exposure of bixafen in soil was assessed at 10 representative field sites in Germany after a period of up to 8 years of use with five different products containing bixafen, followed by annual measurements from 2017 to 2019. The measured exposure concentrations were compared with modeled predicted environmental concentrations in soil (PECsoil) that are derived in the context of the European risk assessment of plant protection products. It was shown that the model assumptions, in particular the kinetic parameters describing the background accumulation, provided a conservative description of the observed residue data. This demonstrates that the exposure modeling tools are adequate for use in soil risk assessment. Laboratory and field ecotoxicological studies were performed to provide a comprehensive risk assessment on the long-term use of bixafen-based fungicides in cereals. While a laboratory reproduction study with the earthworm Eisenia fetida indicated a potential risk at the Tier 1 risk assessment for the end-use product Skyway XPro® , a 2.5-year field study showed no unacceptable long-term effects on natural earthworm populations. The exposure in this study exceeded the maximum recommended field rate of Skyway XPro® by a factor of 3 and the maximum measured bixafen concentrations from exposure monitoring study by a factor of 5.2. Hence, an acceptable long-term risk of bixafen-based cereal fungicides is concluded for earthworms. Integr Environ Assess Manag 2022;18:734-747. © 2021 Bayer AG. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

SDHI Fungicide Toxicity and Associated Adverse Outcome Pathways: What Can Zebrafish Tell Us?

Succinate dehydrogenase inhibitor (SDHI) fungicides are increasingly used in agriculture to combat molds and fungi, two major threats to both food supply and public health. However, the essential requirement for the succinate dehydrogenase (SDH) complex-the molecular target of SDHIs-in energy metabolism for almost all extant eukaryotes and the lack of species specificity of these fungicides raise concerns about their toxicity toward off-target organisms and, more generally, toward the environment. Herein we review the current knowledge on the toxicity toward zebrafish (Brachydanio rerio) of nine commonly used SDHI fungicides: bixafen, boscalid, fluxapyroxad, flutolanil, isoflucypram, isopyrazam, penthiopyrad, sedaxane, and thifluzamide. The results indicate that these SDHIs cause multiple adverse effects in embryos, larvae/juveniles, and/or adults, sometimes at developmentally relevant concentrations. Adverse effects include developmental toxicity, cardiovascular abnormalities, liver and kidney damage, oxidative stress, energy deficits, changes in metabolism, microcephaly, axon growth defects, apoptosis, and transcriptome changes, suggesting that glycometabolism deficit, oxidative stress, and apoptosis are critical in the toxicity of most of these SDHIs. However, other adverse outcome pathways, possibly involving unsuspected molecular targets, are also suggested. Lastly, we note that because of their recent arrival on the market, the number of studies addressing the toxicity of these compounds is still scant, emphasizing the need to further investigate the toxicity of all SDHIs currently used and to identify their adverse effects and associated modes of action, both alone and in combination with other pesticides.

The Short-Term Exposure to SDHI Fungicides Boscalid and Bixafen Induces a Mitochondrial Dysfunction in Selective Human Cell Lines.

Fungicides are used to suppress the growth of fungi for crop protection. The most widely used fungicides are succinate dehydrogenase inhibitors (SDHIs) that act by blocking succinate dehydrogenase, the complex II of the mitochondrial electron transport chain. As recent reports suggested that SDHI-fungicides could not be selective for their fungi targets, we tested the mitochondrial function of human cells (Peripheral Blood Mononuclear Cells or PBMCs, HepG2 liver cells, and BJ-fibroblasts) after exposure for a short time to Boscalid and Bixafen, the two most used SDHIs. Electron Paramagnetic Resonance (EPR) spectroscopy was used to assess the oxygen consumption rate (OCR) and the level of mitochondrial superoxide radical. The OCR was significantly decreased in the three cell lines after exposure to both SDHIs. The level of mitochondrial superoxide increased in HepG2 after Boscalid and Bixafen exposure. In BJ-fibroblasts, mitochondrial superoxide was increased after Bixafen exposure, but not after Boscalid. No significant increase in mitochondrial superoxide was observed in PBMCs. Flow cytometry revealed an increase in the number of early apoptotic cells in HepG2 exposed to both SDHIs, but not in PBMCs and BJ-fibroblasts, results consistent with the high level of mitochondrial superoxide found in HepG2 cells after exposure. In conclusion, short-term exposure to Boscalid and Bixafen induces a mitochondrial dysfunction in human cells.

Bixafen causes cardiac toxicity in zebrafish (Danio rerio) embryos.

Bixafen (BIX) is a succinate dehydrogenase inhibitor (SDHI)-class fungicide that is used to control crop diseases. However, data on the toxicity of BIX to zebrafish are limited. Here, zebrafish embryos were exposed to 0.1, 0.3, and 0.9 µM BIX. After BIX exposure, zebrafish embryos exhibited cardiac dysplasia and dysfunction, including pericardial edema, reduced heart rate, and drastically decreased erythrocytes in the cardiac area; the severity of these negative effects increased with BIX concentration and the duration of BIX exposure. In addition, the transcription levels of erythropoiesis-related genes decreased significantly in BIX-treated embryos, as compared to untreated control embryos. Similarly, compared with the control, key genes responsible for cardiac development (myh6, nkx2.5, and myh7) also exhibited dysregulated expression patterns in response to BIX treatment, suggesting that BIX might specifically affect cardiac development. Finally, cell apoptosis was induced in embryos after BIX treatment. In combination, our results suggested that exposure to BIX induced cardiac toxicity in zebrafish. These data will be valuable for future evaluations of the environmental risks of BIX.

Bixafen exposure induces developmental toxicity in zebrafish (Danio rerio) embryos.

Bixafen (BIX), a new generation succinate dehydrogenase inhibitor (SDHI) fungicide commonly used in agriculture, is regarded as a potential aquatic pollutant because of its lethal and teratogenic effects on Xenopus tropicalis embryos. To evaluate the threat of BIX to aquatic environments, information concerning BIX's embryonic toxicity to aquatic organisms (especially fish) is important, yet such information remains scarce. The present study aimed to fill this knowledge gap by employing zebrafish embryos as model animals in exposure to 0.1, 0.3 and 0.9 µM BIX. Our results showed that BIX caused severe developmental abnormalities (hypopigmentation, tail deformity, spinal curvature and yolk sac absorption anomaly) and hatching delay in zebrafish embryos. The expression levels of early embryogenesis-related genes (gh, crx, sox2 and neuroD) were downregulated after BIX exposure, except for nkx2.4b, which was upregulated. Furthermore, transcriptome sequencing analysis showed that all the downregulated differentially expressed genes were enriched in cell cycle processes. Taken together, these results demonstrated that BIX has strong developmental toxicity to zebrafish that may be due to the downregulated expression of genes involved in embryonic development. These findings provide valuable reference for evaluating BIX's potential adverse effects on aquatic ecosystems.