Anaphalis margaritacea ethanol extract exhibits potent anti-Trichinella spiralis activity via mitochondrial dysfunction and host tissue protection.

Trichinellosis is a food-borne zoonosis caused by Trichinella spiralis, for which effective treatment against the muscle larvae stage remains limited. To identify new anti-T. spiralis agents from medicinal plants, 30 extracts with reported or presumed antiparasitic relevance were screened in vitro. Among them, the 80% ethanol extract of Anaphalis margaritacea (AMEE) showed the strongest larvicidal activity, causing complete muscle larvae mortality within 72 h at 100 μg/mL and exhibiting a clear dose-dependent effect. Preliminary chemical characterization by UPLC-Q-TOF-MS/MS led to the tentative identification of eight major constituents, mainly organic acids and flavonoids. In a murine model of T. spiralis infection, oral administration of AMEE reduced muscle larvae burden by up to 59.7% in a dose-dependent manner. AMEE treatment also alleviated muscle fibre damage and inflammatory infiltration in infected muscle, accompanied by reduced levels of IL-6, IL-13, and TGF-β and decreased mRNA expression of Nqo1 and Sod2. To explore the basis of its antiparasitic activity, integrated proteomics, mitochondrial enzyme activity assays, and transmission electron microscopy were performed. AMEE markedly suppressed the activities of mitochondrial respiratory complexes I, II, and IV, reduced complex V activity, and induced mitochondrial swelling and cristae disruption in muscle larvae. These findings show that AMEE exerts significant anti-T. spiralis activity in vitro and in vivo, with evidence consistent with mitochondrial dysfunction in muscle larvae and attenuation of infection-associated muscle pathology.