FIG 5.

Entamoeba muris shows reliable excystation in vitro when treated with upper gastrointestinal tract components. Fecal cysts were purified by sucrose density gradient and then acid washed (0.1 M HCl). Cysts were inoculated into excystation conditions (1% bovine bile, 80 mM sodium bicarbonate, or a combination of both), then incubated for 24 h at room temperature. (A) Cysts were scored from 0 to 3, where 0 represented an intact cyst and 3 is an empty chitin shell. Chitin (Calcofluor White), Jacob2 (1A4 antibody [17]), and nuclei (Syto11). Scale bar represents 10 μm. (B) Excystation rates (score ≥ 1) were quantified under these conditions. Significance was determined using a two-tailed t test. Only significant pairwise comparisons are shown; Bile (P = 0.004) and, NaHCO₃ + Bile (P = 0.0064). Each dot represents a biological replicate (n = 3 independent experiments), black horizontal line is the average of the three biological replicates.