The aim of this study was to investigate the effect of dietary probiotic inclusion level on mucin composition (monosaccharide ratio), mucosal morphometry, mucus layer staining intensity, and mucus layer thickness along the broiler intestinal tract. One-day-old male Cobb broilers were administered maize-soybean meal basal (BD) diets for 42 d and depending on the feed additive used, broilers were allocated into the following 5 experimental treatments: control C (BD, no additive), treatment P1 (10(8) colony forming units of probiotic/kg of BD), treatment P2 (10(9) cfu of probiotic/kg of BD), treatment P3 (10(10) cfu of probiotic/kg of BD), and treatment A (2.5 mg avilamycin/kg of BD). Intestinal samples from duodenum, ileum, and cecum of 14- and 42-d-old broilers were collected and analyzed. Mannose (Man) decreased linearly with increasing probiotic level in duodenum (P=0.015) and ileum (P=0.042) of 14-d-old broilers. N-Acetyl-glucosamine and galactose decreased linearly (P=0.012 and P=0.001, respectively), while fucose increased linearly (P<0.001) with increasing probiotic feed inclusion level in 42-d-old broiler cecum, with treatment A not differing from treatment C (P≥0.05). Cecal villus height and crypt depth increased linearly (P=0.016 and P=0.003, respectively) with probiotic inclusion level, with treatment A having higher (P≤0.05) values only from treatment C. Mucus layer thickness increased linearly with probiotic inclusion level in duodenum at 14 d and 42 d (P=0.007 and P=0.030, respectively). Finally, mucus layer staining intensity was influenced (P<0.001) by villus fragment (i.e., tip, midsection, and base) but not from the treatment, age, and intestinal segment examined. As a conclusion, this study provides evidence that probiotic inclusion level affects intestinal mucin monosaccharide composition, mucus layer thickness, and intestinal morphology in broilers.