Development Of Zebrafish Model For Comprehensive Assessment Of Endocrine Disruption Based On Hormone Biomarkers And Application In Assessment For Three Fungicides

With the development of industry and society,more endocrine disrupting compounds?EDCs?are introduced to our life.These compounds produce endocrine disrupting effects by interfering biological activities including growth,development,reproduction,metabolism,and so on.At present,most of studies involving the assessment of EDCs focused on only one or few types of endocrine effects.In order to fully understand and evaluate the endocrine disruption of EDCs,it is urgent to develop a new model to comprehensively evaluate the multiple endocrine effects of EDCs.In the study,a zebrafish model was developed to evaluate the comprehensive endocrine disruption of EDCs by monitoring 23 hormones to indicate the effects on different endocrine systems including epinephrine,thyroxine,and pituitary systems with UPLC-MS/MS.The new approach can effectively remove matrix interference and reach to satisfied recoveries,accuracy and precision for these hormone biomarkers.The matrix effects are mainly between 67.3%and 132.1%.The relative recoveries mainly range from 73.4%to 110.2%.The errors of method are less than13.8%,and the relative standard deviations range from 0.3%to 20.0%.The endocrine disrupting effects of three triazole fungicides including propiconazole,tebuconazole and myclobutanil were then studied with the new model.The results indicated that the fungicides showed hormone-specific disruption on the23 biomarkers.Generally,their effects on peptide hormones and fatty acid derivative hormones were greater than the other hormones including thyroid hormones.Myclobutanil possessed the greatest potential of disruption among the three fungicides.Similar trends of effects were observed for some of the hormones such as cortisol?F?and thromboxane B2?TX B2?among the fungicide-exposed groups.But inconsistent trends were found for the other hormones such as norepinephrine?NE?and melatonin?MT?.Combined effects of endocrine disruption were also evaluated for the three fungicides at exposure level of 100 ng L^-1.Similar gender-specific trends of effects were observed on most of the hormones in the combined exposure compared to the exposure of individual fungicide.Combined exposure also suggested antagonistic effects on most of the hormones such as aldosterone?ALDO?and synergistic effect on a few hormones such as prostaglandin D2?PG D₂?.Two comprehensive evaluation methods,IBR?Integrated Biomarker Response?and Tox Pi?Toxicological Priority Index?,were employed to analyze the endocrine disruption of the three fungicides at three levels of hormone biomarker,endocrine system and comprehensive index.Tox Pi was also optimized.Good consistence between the two methods and gender-specific difference of endocrine disruption were observed for the fungicides.The results indicated a greater endocrine disruption at low concentration and a dose-response curve of reverse“U”.In combined exposure at100 ng·L^-1,the endocrine effect of myclobutanil is larger than that of tebuconazole and propiconazole.Combined exposure generally lead to antagonistic effect.The new model developed in the study can comprehensively evaluate endocrine disruption of EDCs,achieve overall and intuitive assessment for EDCs,and provide an alternative approach for screening of EDCs and evaluating environmental quality based on endocrine disrupting effects.