Structural Characterization of Toxoplasma gondii Brain Cysts in a Model of Reactivated Toxoplasmosis Using Computational Image Analysis

Toxoplasma gondii is an obligate intracellular parasite existing in three infectious life stages-tachyzoites, bradyzoites, and sporozoites. Rupture of tissue cysts and re-conversion of bradyzoites to tachyzoites leads to reactivated toxoplasmosis (RT) in an immunocompromised host. The aim of this study was to apply ImageJ software for analysis of T. gondii brain cysts obtained from a newly established in vivo model of RT. Mice chronically infected with T. gondii (BGD1 and BGD26 strains) were treated with cyclophosphamide and hydrocortisone (experimental group-EG) or left untreated as infection controls (ICs). RT in mice was confirmed by qPCR (PCR+); mice remaining chronically infected were PCR-. A total of 90 images of cysts were analyzed for fractal dimension (FD), lacunarity (L), diameter (D), circularity (C), and packing density (PD). Circularity was significantly higher in PCR+ compared to IC mice (p < 0.05 for BGD1, p < 0.001 for the BGD26 strain). A significant negative correlation between D and PD was observed only in IC for the BGD1 strain (rho = -0.384, p = 0.048), while fractal parameters were stable. Significantly higher D, C, and PD and lower lacunarity, L, were noticed in the BGD1 compared to the more aggressive BGD26 strain. In conclusion, these results demonstrate the complexity of structural alterations of T. gondii cysts in an immunocompromised host and emphasize the application potential of ImageJ in the experimental models of toxoplasmosis.