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. 2021 Feb 4;10:632556. doi: 10.3389/fcimb.2020.632556

Figure 2.

Figure 2

The presence of immature second generation schizonts in germ-free chickens at day 3.5 post-infection suggests a delay in the development of the parasite. (A) Gametocyte gene expressions are delayed in germ-free chickens. Sporulated oocysts were orally inoculated to germ-free and conventional chickens (500,000 for 2 days and 10,000 for 3.5, 5.5, and 7 days of infection). Gametocyte gene expressions were detected by RT-qPCR on caecal tissues. The expression of microgamete and macrogamete specific genes (Etfoa1 and Etgam56) were determined by RT-qPCR and expressed relative to E. tenella housekeeping genes (Et18S and Etprofilin) at day 2, 3.5, 5.5, and 7 post-infection using the 2-ΔCt formula: Ct Et gametocyte specific gene – Ct Et housekeeping genes. Medians are represented (One-Way ANOVA, Dunn’s multiple comparisons test; n ≥ 3; *P< 0.05; ***P <0.001 between germ-free and conventional chickens). (B) Representative histological images showing a delay in parasite development in germ-free chickens. Histological analysis was performed on tissue sections stained with Masson-Goldner Trichrome. Scale = 20 µm; Schz, schizont; Ma, macrogamete; Oo, oocyst.