Giardia duodenalis triggered neutrophil extracellular traps in goats

Giardia duodenalis is a globally distributed zoonotic parasite primarily transmitted through the fecal-oral route, infecting various vertebrates, and the infection of which is prevalent in goats. Immune cells play a crucial role in pathogens invasion, and neutrophil extracellular traps (NETs) released by neutrophils serve as a non-specific defense mechanism against pathogens including parasites. In this study, we investigated the characteristics, components, and molecular mechanisms of goat NETs upon stimulation with G. duodenalis trophozoites. This study demonstrates that G. duodenalis trigger dose-dependent NETs formation in goat neutrophils, composed of DNA, citrullinated histone H3 (CitH3), and neutrophil elastase (NE). Reactive oxygen species (ROS) accumulation synchronizes with NETosis during G. duodenalis infection. Inhibitor experiments confirmed that G. duodenalis-induced NETs and ROS production depend on TLR2/4 signaling and require NADPH oxidase (NOX), ERK_1/2, and p38 MAPK activation. This work identifies TLR2/4, NOX, ERK_1/2, and p38 MAPK pathways as key regulators of NETs/ROS coordination during G. duodenalis infection, providing the first evidence of G. duodenalis-triggered NETs in goats. The findings highlight NETs as critical components of anti-G. duodenalis immunity and suggest potential for NETs-targeted therapeutic strategies.