Metabolic Profile Associated With Encystation in <i>Acanthamoeba</i>

The genus Acanthamoeba includes widespread protozoa that can cause severe infections in humans. Their ability to form resistant cysts within infected tissues complicates treatment, making it essential to understand the encystation process for developing effective therapeutic strategies. This study utilized untargeted metabolomics (GC-MS) to analyze metabolic changes during the encystation of an Acanthamoeba strain in Neff's encystation saline. We conducted metabolite analysis at three stages of differentiation: the trophozoite-dominant phase (0 h), the pre-cyst-dominant phase (24 h), and the cyst-dominant phase (72 h). The results indicated a global metabolic downregulation during encystation, which is consistent with a state of dormancy. Components of the cyst wall such as cellobiose and lactose accumulated in the final phase. Arbutin and canavanine were annotated for the first time in Acanthamoeba. Encystation also led to changes in pathways related to glycine, serine, and threonine metabolism and biosynthesis of aminoacyl-tRNA. This study uncovered previously unknown metabolites and metabolic pathways at distinct stages of Acanthamoeba development.