Chronic chlorothalonil exposure inhibits locomotion and interferes with the gut-liver axis in Pelophylax nigromaculatus tadpoles

Chlorothalonil is a widely used fungicide that has a negative effect on individual movement, but its impact pathway needs further refinement. Here, the effects of exposure to chlorothalonil on the locomotion behavior of Pelophylax nigromaculatus tadpoles (GS23) were measured at three different levels (0 µg/L, 10 µg/L, and 50 µg/L), and the possible pathways of its effects were analyzed from the gut-liver axis. Chlorothalonil exposure levels of 10 µg/L and 50 µg/L significantly reduced the average speed of P. nigromaculatus tadpoles by 26% and 32.7%, respectively, and significantly decreased the locomotor frequency by 27.1% and 58.6%, respectively. Gut microbiota analysis revealed chlorothalonil exposure significantly increased the abundance of Firmicutes, while significantly decreased the abundance of Actinobacteriota, Pseudomonas, and Rhodococcus. Metabolomics analysis identified that chlorothalonil treatment changed amino acid-related metabolism pathways in the gut and liver and altered the glycerophospholipid metabolism pathway in the liver. This study indicated that chlorothalonil can affect individual locomotor abilities and interfering with the gut-liver axis of aquatic animals. These findings establish that chlorothalonil compromises aquatic organism motility through a multi-target mechanism involving gut microbiota modulation, amino acid metabolic interference, and hepatic lipid pathway disruption.