Dot-plot graphs of N. caninum burdens in cotyledons. N. caninum burden in cotyledons from intravenously challenged dams (A) and from IV and SC challenged animals with 104 Nc-Spain7 tachyzoites (B). Each dot represents individual values of parasite burden (number of parasites per mg of host tissue), and medians are represented as horizontal lines. Considering that the N. caninum detection limit by real-time PCR is 0.1 parasites, negative samples (0 parasites) were represented on the log scale as < 0.1 (i.e., 10−2). Parasite burdens were analysed using the non-parametric Kruskal–Wallis test followed by the Dunn’s test for comparisons between groups, as well as the Mann–Whitney test for pairwise comparisons. (**) indicates P < 0.01, (***) P < 0.001, and (ρ) indicates a trend towards significant differences among infected groups in each tissue.