Analysis of the Therapeutic Effect of Gallic Acid Against Ichthyophthiriasis in Coreius guichenoti and Its Liver Metabolome
HUANG Lei1,2, PENG Xian-Qi2, BU Xia-Lian2, ZHAO Yu1, LIU Yu-Long2, CHEN Jing2, NIU Chen2, ZHU Jian1, QU Huan-Tao1,*, YAO Jia-Yun1,2,*
1 Hubei Key Laboratory of Three Gorges Project for Conservation of Fishes/Chinese Sturgeon Research Institute, China Three Gorges Corporation, Yichang 443100, China; 2 Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture and Rural Affairs/Key Laboratory of Fish Health and Nutrition of Zhejiang Province/Key Laboratory of Fishery Environment and Aquatic Product Quality and Safety of Huzhou City, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China
Abstract：The largemouth bronze gudgeon (Coreius guichenoti), a rare endemic fish species in the upper Yangtze River, faces critical survival challenges due to ichthyophthiriasis. In order to investigate the therapeutic efficacy of plant-derived gallic acid (GA) against Ichthyophthirius multifiliis infection and elucidate its molecular mechanisms, this study systematically evaluated the therapeutic efficacy, safety profile, and underlying mechanisms of action of GA through in vitro and in vivo antiparasitic assays, acute toxicity tests, and hepatic non-targeted metabolomics analysis. In vitro assays demonstrated that GA exhibited concentration-dependent antiparasitic effects on I. multifiliis. After 4 h of treatment with 50 mg/L GA, the mortality of trophonts and theronts reached 100.0%, while 20 mg/L GA resulted in 63.3% and 80.3% mortality, respectively. Microscopic observations revealed pathological alterations in parasites, including cell membrane disruption, cytoplasmic leakage, and mitotic arrest in the parasites following GA treatment. In vivo trials showed that 50 and 20 mg/L GA reduced fish mortality from 100% (control) to 2.3% and 20.0% within 48 h, respectively. Concurrently, parasite mortality reached 54.3% and 21.3%. Moreover, GA treatment alleviated clinical symptoms, including white spots, excessive mucus secretion, and respiratory distress. Acute toxicity tests determined that the safe concentration (SC) of GA for C. guichenoti was 64.6 mg/L, with 96 h half-lethal concentration (LC50) values of 245.8 mg/L. Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis revealed that 50 mg/L GA significantly reshaped the hepatic metabolic network in I. multifiliis-infected fish: (1) downregulating arachidonic acid metabolism, VEGF signaling, and Th17 cell differentiation pathways to suppress pro-inflammatory mediators; (2) upregulating endogenous antioxidants (e.g., glutathione and taurine) while activating FoxO signaling and lysosomal pathways to enhance reactive oxygen species scavenging; (3) optimizing mitochondrial energy metabolism and membrane homeostasis by enriching pathways such as oxidative phosphorylation, glycerophospholipid metabolism, and the malate-aspartate shuttle. These findings demonstrated that 50 mg/L GA effectively controled ichthyophthiriasis via a synergistic mechanism involving anti-inflammatory regulation, antioxidant defense activation, and metabolic reprogramming, thereby restoring hepatic metabolic homeostasis. This study provides novel strategies and a candidate agent for the environmentally friendly control of ichthyophthiriasis in fish.
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