The widespread application of pesticides in agriculture has raised increasing concerns regarding their ecological impact, particularly in aquatic environments. Among these, α-cypermethrin, a highly active isomeric form of cypermethrin, has been extensively used due to its potent insecticidal efficacy and low mammalian toxicity. However, its toxicity to non-target aquatic organisms remains insufficiently understood at the metabolic level. In this study, a targeted metabolomics approach was employed to investigate the biochemical effects of α-cypermethrin in adult zebrafish. Acute toxicity was first determined to establish sublethal exposure concentrations (0.15 µg/L and 1.5 µg/L), followed by a 48 h exposure under a controlled flow-through system. GC-MS/MS-based analysis quantified 395 metabolites, and multivariate statistical models (principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA)) revealed clear dose-dependent metabolic alterations at two time points. Pathway analysis identified disruptions in glycolysis, glycerolipid metabolism, amino acid turnover, and glutathione pathways. Notably, glutamate depletion and associated reductions in GABA (4-Aminobutanoate) and TCA (Tricarboxylic acid) cycle intermediates suggest oxidative stress-induced metabolic bottlenecks. These results provide mechanistic insights into α-cypermethrin-induced toxicity and demonstrate the utility of metabolite-level biomarkers for environmental monitoring. This study contributes to a systems-level understanding of how sublethal pesticide exposure affects vertebrate metabolism, offering a basis for improved ecological risk assessment and pesticide regulation.

Accidental ingestion of toxic mushrooms remains a global public health concern because of the presence of highly potent peptide toxins such as α‐amanitin, β‐amanitin, and phalloidin. These compounds exhibit strong hepatotoxicity and can lead to acute liver failure and death. However, their rapid detection in biological and food matrices remains analytically challenging. Existing methods often require extensive sample preparation and are not suitable for urgent diagnostic applications. This study presents the development and validation of a rapid and sensitive analytical method for the simultaneous quantitation of α‐amanitin, β‐amanitin, and phalloidin in poisonous mushrooms and human serum. Among several preparation strategies evaluated, a method following direct extraction with 1% formic acid in methanol was selected for its speed, simplicity, and effectiveness in minimizing matrix interference. The method demonstrated excellent linearity (r² ≥ 0.99), low quantitation limits (10–50 ng/mL), and satisfactory recovery (72%–117%) and precision (RSD ≤ 19%) in both food and biological matrices. When applied to field‐collected Amanita virosa, α‐amanitin and β‐amanitin were detected at 39 and 145 mg/kg, respectively, whereas no toxins were found in Amanita volvata. These findings demonstrate that the established method enables rapid and reliable detection of lethal peptide toxins with minimal sample preparation. The protocol is suitable for forensic investigations, clinical toxicology, and food safety monitoring. Its applicability in emergency settings underscores its potential as a practical tool for improving public health responses to mushroom poisoning incidents.

As climate-driven expansion of tick populations intensifies the use of neonicotinoid-based veterinary products worldwide, the extent of internal exposure in companion animals and the associated health risks remain poorly understood. This study measured the concentrations of nine neonicotinoids and eight of their metabolites in urine samples from 47 dogs. The results showed that neonicotinoids and their metabolites were detected in 97% of the samples. The most frequently found compounds were clothianidin-desmethyl (51%), imidacloprid-olefin (43%), and imidacloprid (34%). Imidacloprid-olefin showed the highest concentration at 383 ng/mL, followed by 5-hydroxy-imidacloprid at 44 ng/mL. Urinary neonicotinoid levels did not differ significantly by sex, body size, or age, indicating that chemical use patterns may be a more influential factor in exposure than individual traits. The highest chronic daily intake (CDI) was for the metabolite imidacloprid-olefin at 28 μg/kg/day, indicating greater exposure to imidacloprid, followed by N-desmethyl-thiamethoxam at 1.84 μg/kg/day. While median hazard quotient (HQ) values remained below 1, the maximum HQ for imidacloprid-olefin reached 0.47, a level that warrants caution given uncertainties in human-derived reference doses. The widespread co-detection of multiple compounds indicates continuous chronic exposure and raises concern over uncharacterized mixture effects. Their extensive use and excretion into the environment may contribute to secondary exposure among cohabitants and potentially affect non-target organisms within the broader urban ecosystem.