Fluconazole (FLU) is a widely used antifungal agent frequently detected in surface waters because of its extensive use in medicine, agriculture, and personal care products.Despite concerns about its persistence and developmental toxicity in aquatic species, its effects on amphibians remain poorly understood. This study aimed to assess the developmental and molecular effects of FLU and its structural core, 1,2,4-triazole (TRI), in amphibian embryos. Xenopus laevis embryos were exposed to FLU or TRI and evaluated for mortality, hatching rate, heart rate, body length, malformation incidence, and changes in gene expression. Even at low micromolar concentrations, both azoles altered the expression of Wnt- and BMP-associated genes, indicating disruption of these signaling pathways. At higher micromolar concentrations, these molecular changes were accompanied by early signs of developmental abnormalities, which intensified at the highest doses. Observed phenotypes included reduced head size, altered skin pigmentation, prolonged body length, changes in heart rate, and mild digestive tract malformations. These findings demonstrate that even the core structural motif TRI can disrupt key developmental signaling pathways in vertebrate embryos, underscoring the need for closer monitoring of azole compounds in aquatic environments. Given the fundamental role of these pathways in vertebrate development, the results raise concerns about potential risks from long-term or prenatal exposure to azoles, in both environmental and clinical contexts.

Department of Animal Protection and Welfare and Veterinary Public Health University of Veterinary Sciences Brno Czech Republic

Department of Experimental Biology Masaryk University Brno Czech Republic

Department of Infectious Diseases and Preventive Medicine Veterinary Research Institute Brno Czech Republic

Zobrazit více v PubMed

Chakraborty, A. et al. Pharmaceuticals and personal care products as emerging environmental contaminants: Prevalence, toxicity, and remedial approaches. DOI

Jørgensen, L. N. & Heick, T. M. Azole use in agriculture, horticulture, and wood preservation—Is it indispensable?. PubMed DOI PMC

Sheehan, D. J., Hitchcock, C. A. & Sibley, C. M. Current and emerging azole antifungal agents. PubMed DOI PMC

Lee, Y., Robbins, N. & Cowen, L. E. Molecular mechanisms governing antifungal drug resistance. PubMed DOI PMC

Maertens, J. A. History of the development of azole derivatives. PubMed DOI

Gupta, O., Pradhan, T. & Chawla, G. An updated review on diverse range of biological activities of 1,2,4-triazole derivatives: Insight into structure activity relationship. DOI

Bhagat, J., Singh, N., Nishimura, N. & Shimada, Y. A comprehensive review on environmental toxicity of azole compounds to fish. PubMed DOI

Kahle, M., Buerge, I. J., Hauser, A., Müller, M. D. & Poiger, T. Azole fungicides: Occurrence and fate in wastewater and surface waters. PubMed DOI

Lu, Z.-J. et al. The azole biocide climbazole induces oxidative stress, inflammation, and apoptosis in fish gut. PubMed DOI

Płatkiewicz, J., Frankowski, R., Cieślak, A., Grześkowiak, T. & Zgoła-Grześkowiak, A. Long-term study of azoles in surface water and treated wastewater. PubMed DOI

Subramanian, A. et al. Comparative risk assessment studies estimating the hazard posed by long-term consumption of PPCPs in river water. PubMed DOI

Halešová, T., Konečná, J., Václavíková, M., Karásek, P., Nováková, E., 2022. Výskyt pesticidních látek v řece Punkvě. VTEI 64, 29. 10.46555/VTEI.2021.12.001

Iancu, V.-I. et al. Occurrence and distribution of azole antifungal agents in eight urban Romanian waste water treatment plants. PubMed DOI

Liu, W.-R. et al. Biocides in the Yangtze River of China: Spatiotemporal distribution, mass load and risk assessment. PubMed DOI

Wick, A., Fink, G. & Ternes, T. A. Comparison of electrospray ionization and atmospheric pressure chemical ionization for multi-residue analysis of biocides, UV-filters and benzothiazoles in aqueous matrices and activated sludge by liquid chromatography–tandem mass spectrometry. PubMed DOI

Wilkinson, J. L. et al. Pharmaceutical pollution of the world’s rivers. PubMed DOI PMC

Chen, Z.-F. et al. Photodegradation of the azole fungicide fluconazole in aqueous solution under UV-254: Kinetics, mechanistic investigations and toxicity evaluation. PubMed DOI

Fahy, W. D., Zhang, Z., Wang, S., Li, L. & Mabury, S. A. Environmental fate of the azole fungicide fluconazole and its persistent and mobile transformation product 1,2,4-triazole. PubMed DOI

Barenys, M., Molins, A., Amorós-Galicia, L., Flick, B. & Gómez-Catalán, J. Implementation of a functional endpoint to the zebrafish embryotoxicity test to evaluate craniofacial abnormalities. PubMed DOI

De Robertis, E. M. & Gurdon, J. B. A brief history of PubMed DOI

Gao, J. & Shen, W. Xenopus in revealing developmental toxicity and modeling human diseases. PubMed DOI

Mouche, I., Malésic, L. & Gillardeaux, O. FETAX Assay for Evaluation of Developmental Toxicity. In PubMed

Mouche, I., Malesic, L. & Gillardeaux, O. FETAX Assay for Evaluation of Developmental Toxicity. In PubMed

Rodrigues, M. D. S., Toledo, V. P. B. & Nóbrega, R. H. Effects of maternal stress on the development of the somatotropic axis during the larval and juvenile stages in Zebrafish ( DOI

Senarath Pathirajage, K. & Rajapaksa, G. Long-term exposure to environmentally relevant Bisphenol-A levels affects growth, swimming, condition factor, sex ratio and histology of juvenile zebrafish. PubMed DOI PMC

Hu, Y. & Bajorath, J. Structural and activity profile relationships between drug scaffolds. PubMed DOI PMC

Janowski, M., Demchuk, O. M. & Wujec, M. Fluconazole analogs and derivatives: An overview of synthesis, chemical transformations, and biological activity. PubMed DOI PMC

Tiboni, G. M. & Giampietro, F. Murine teratology of fluconazole: Evaluation of developmental phase specificity and dose dependence. PubMed DOI

Sattar, R.Z. Toxicological effects of Fluconazole on chick embryo: A literature review. JCRMHS 4. (2023) 10.55920/JCRMHS.2023.04.001155

Bhagat, J., Zang, L., Nakayama, H., Nishimura, N. & Shimada, Y. Effects of nanoplastic on toxicity of azole fungicides (ketoconazole and fluconazole) in zebrafish embryos. PubMed DOI

Wu, H., Zhao, G., Feng, W., Yang, C. & Jiang, Y. Fluconazole induces cardiovascular toxicity in zebrafish by promoting oxidative stress, apoptosis, and disruption of key developmental genes. PubMed DOI

Iuga, A., Lerner, E., Shedd, T. R. & Van Der Schalie, W. H. Rapid responses of a melanophore cell line to chemical contaminants in water. PubMed DOI

Dickinson, A. J. G. Using frogs faces to dissect the mechanisms underlying human orofacial defects. PubMed DOI PMC

Anderson, R. M., Lawrence, A. R., Stottmann, R. W., Bachiller, D. & Klingensmith, J. Chordin and noggin promote organizing centers of forebrain development in the mouse. PubMed DOI

Piccolo, S. et al. Cleavage of chordin by xolloid metalloprotease suggests a role for proteolytic processing in the regulation of spemann organizer activity. PubMed DOI PMC

Clevers, H. Wnt/β-catenin signaling in development and disease. PubMed DOI

Hikasa, H. & Sokol, S. Y. Wnt signaling in vertebrate axis specification. PubMed DOI PMC

McMahon, J. A. et al. Noggin-mediated antagonism of BMP signaling is required for growth and patterning of the neural tube and somite. PubMed DOI PMC

Stottmann, R. W., Anderson, R. M. & Klingensmith, J. The BMP antagonists chordin and noggin have essential but redundant roles in mouse mandibular outgrowth. PubMed DOI

Kelsh, R. N., Harris, M. L., Colanesi, S. & Erickson, C. A. Stripes and belly-spots—A review of pigment cell morphogenesis in vertebrates. PubMed DOI PMC

Raible, D. W. & Ragland, J. W. Reiterated Wnt and BMP signals in neural crest development. PubMed DOI

Arnold, S. J. et al. Brachyury is a target gene of the Wnt/β-catenin signaling pathway. PubMed DOI

Tzahor, E. Wnt/β-catenin signaling and cardiogenesis: Timing does matter. PubMed DOI

Foulquier, S. et al. WNT signaling in cardiac and vascular disease. PubMed DOI PMC

Li, D., Sun, J. & Zhong, T. P. Wnt signaling in heart development and regeneration. PubMed DOI

Tian, Y., Cohen, E. D. & Morrisey, E. E. The importance of Wnt signaling in cardiovascular development. PubMed DOI PMC

Ben Othmène, Y. et al. Triazole fungicide tebuconazole induces apoptosis through ROS-mediated endoplasmic reticulum stress pathway. PubMed DOI

Li, S. et al. Tebuconazole induced oxidative stress related hepatotoxicity in adult and larval zebrafish ( PubMed DOI

Yang, J. et al. Effects of tebuconazole on cytochrome P450 enzymes, oxidative stress, and endocrine disruption in male rats. PubMed DOI

Lum, H. & Roebuck, K. A. Oxidant stress and endothelial cell dysfunction. PubMed DOI

Scallan, J., Huxley, V. H. & Korthuis, R. J. Capillary fluid exchange: Regulation, functions, and pathology. PubMed DOI

Dong, B. A comprehensive review on toxicological mechanisms and transformation products of tebuconazole: Insights on pesticide management. PubMed DOI

Domingos, E. L. et al. Comparative efficacy and safety of systemic antifungal agents for candidemia: A systematic review with network meta-analysis and multicriteria acceptability analyses. PubMed DOI

Katiraee, F., Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran, Teifoori, F., Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy and laboratory of Parasitology and Allergy, Lascaray Research Center, University of the Basque Country, Vitoria, Spain, Soltani, M., Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran,. Emergence of Azoles Resistance Candida species in Iranian AIDS defined patients with oropharyngeal candidiasis. mazu-cmm 1, 11–16. (2015) 10.18869/acadpub.cmm.1.3.11

Ostrosky-Zeichner, L. Combination antifungal therapy: A critical review of the evidence. PubMed DOI

Ramli, A. H. et al. Antifungal activity of synthetic xanthenone against fluconazole-resistant PubMed DOI

Sanyaolu, A. et al. PubMed DOI PMC

Worku, M. & Girma, F. PubMed DOI PMC

Shin, J., Kim, J.-E., Lee, Y.-W. & Son, H. Fungal cytochrome P450s and the P450 complement (CYPome) of Fusarium graminearum. PubMed DOI PMC

Song, J., Zhang, S. & Lu, L. Fungal cytochrome P450 protein Cyp51: What we can learn from its evolution, regulons and Cyp51-based azole resistance. DOI

Zhang, X., Guo, J., Cheng, F. & Li, S. Cytochrome P450 enzymes in fungal natural product biosynthesis. PubMed DOI

Guengerich, F. P., Waterman, M. R. & Egli, M. Recent structural insights into cytochrome P450 function. PubMed DOI PMC

Hannemann, F., Bichet, A., Ewen, K. M. & Bernhardt, R. Cytochrome P450 systems—biological variations of electron transport chains. PubMed DOI

Zahn, N. et al. Normal Table of PubMed DOI PMC