The three treatments were allocated to cages in a completely randomized block (row of cages) design to give 12 replicates per treatment. Birds had free access to treatments and drinking water for 14 d; feed was replenished throughout the day and feed refusals were weighed daily, for determining cage feed intake. On d 5, birds (6 replicates per treatment on the left row of cages) were challenged with 1 mL of Eimeria culture (25,000 oocysts of E. acervulina and 5,000 oocysts of E. maxima) in distilled water suspension via oral gavage and the other 6 replicates (non-challenged control, on the right row of cages) were given equal volume of distilled water. The separation of right and left cages was an effort to minimize cross-contamination of non-challenge cages. Moreover, daily checks and servicing of the birds started with non-challenged birds followed by challenge birds. The Eimeria culture and challenge protocols were provided by Dr. John Barta of Department of Pathobiology, University of Guelph. Body weight and feed intake was monitored during pre- (d 0 to 5) and post- (d 6 to 14) challenge periods for calculation of BWG and FCR. Two birds per cage were Necropsied on d 10 for intestinal tissue samples. Jejunum was immediately located and excised at duodenal loop and 2 cm anterior to Markel diverticulum. Segments (∼3 cm) of mid-jejunum were excised and placed in buffered formalin for histomorphology analysis (Kiarie et al., 2007 (link)). Additional segments of mid-jejunum (∼1 cm) were placed in a 2 mL tube filled with 1.2 mL Ambion RNAlater (Life Technologies Inc., Burlington, ON, Canada). These samples were placed on ice and immediately transported to the lab and stored at −20°C until required for mRNA analysis of digestive enzymes, nutrients transporters, tight junction proteins, and cytokines. Lesion scores in intestinal regions (duodenum, jejunum, ileum, and ceca) were assessed blindly as described by Price et al. (2014 ) using a scale of 0 (none) to 4 (high) (Johnson and Reid, 1970 (link)). Excreta samples for apparent retention (AR) of components and oocyst shedding were collected from d 10 to 13. The excreta samples for oocyst counts were collected, stored at 4°C, and processed in accord with Price et al. (2014 ). The excreta samples for AR of components were frozen at −20°C until required for analyses. All birds were Necropsied by cervical dislocation on d 14 for gastrointestinal weight measurements