Trifloxystrobin (TFS) is a strobilurin fungicide commonly used in agriculture that inhibits mitochondrial respiratory chain complex III in fungi. Given its widespread use, it is crucial to evaluate its toxic effects on aquatic organisms. This study investigated the developmental toxicity and metabolomic changes in zebrafish (Danio rerio) embryos exposed to TFS for up to 96 hours post-fertilization (hpf). TFS exposure significantly reduced the survival and hatching rates of zebrafish embryos in a concentration-dependent manner, with a 96 h LC50 value of 264.9 μg L-1. Morphological analysis revealed several developmental abnormalities, including yolk sac edema, pericardial edema, reduced body length, and severe developmental delay. Metabolomic analysis using GC-TOF-MS demonstrated a clear distinction between control and TFS-treated groups in PLS-DA score plots, indicating significant alterations in metabolic profiles due to TFS exposure. A total of 42 differential metabolites were identified, encompassing organic acids, amino acids, carbohydrates, and fatty acid-related metabolites. Specifically, glutaric acid, glyceric acid, citric acid, and several fatty acid derivatives increased in the TFS-treated group, while oxalic acid, fumaric acid, malic acid, glucose, and fructose decreased. Pathway enrichment analysis highlighted significant disruptions in fatty acid biosynthesis, the TCA cycle, amino acid metabolism, and carbohydrate metabolism pathways. These findings indicate that TFS exposure interferes with multiple metabolic pathways critical for energy metabolism and developmental processes in zebrafish embryos, thereby suggesting potential ecological risks associated with TFS contamination in aquatic environments.

Abstract
1. 서 론
2. 재료 및 방법
    2.1. 시험 물질
    2.2. 제브라피쉬 사육 조건 및 배아 수집
    2.3. TFS의 제브라피쉬 배아에 대한 급성 독성 평가
    2.4. 대사체 분석을 위한 제브라피쉬 배아 시료 준비
    2.5. 대사체 추출
    2.6. GC-TOF-MS를 이용한 대사체 분석
    2.7. 데이터 처리 및 통계 분석
3. 결 과
    3.1. 제브라피쉬 배아에서 TFS 노출에 따른 생존율 및부화율
    3.2. 제브라피쉬 배아에서 TFS의 치사농도
    3.3. TFS 노출에 따른 표현형 및 전장 길이 변화
    3.4. TFS 노출에 따른 제브라피쉬 배아의 대사체 변화
    3.5. TFS 노출에 따른 대사경로 변화 분석
4. 고 찰
    4.1. 제브라피쉬 배아에 대한 TFS의 급성독성
    4.2. TFS 노출에 따른 제브라피쉬 배아의 대사체 변화
5. 결 론
적 요
CRediT authorship contribution statement
Declaration of Competing Interest
사 사
REFERENCES

• 김채은(경북대학교 응용생명과학과) | Chaeeun Kim (Department of Applied Biosciences, Kyungpook National University, Daegu 41566, Republic of Korea)
• 이상만(경북대학교 응용생명과학과) | Sangman Lee (Department of Applied Biosciences, Kyungpook National University, Daegu 41566, Republic of Korea)
• 이성은(경북대학교 응용생명과학과) | Sung-Eun Lee (Department of Applied Biosciences, Kyungpook National University, Daegu 41566, Republic of Korea) Corresponding author