Quantifying Parasites in Animal Tissues

Parasite quantification in tissues was performed following the methods by Santoro et al. [18 (link)]. Animal tissues were collected at 8 days post infection and stored in individual vials at − 20 °C before genomic DNA extraction. For each sample, 1 g of tissue was individually homogenized, and 200 µl of the homogenate was used to extract genomic DNA according to the manufacturer’s protocols (DP304, Tiangen, China). A standard curve was obtained by linear regression analysis of the threshold cycle (Ct) value (y-axis) versus the log of the initial copy number present in each sample dilution (x-axis). PCR efficiency (E) was calculated as E = 10 (1/slope) ⁻¹.