The negative effects of aflatoxins (AFLA) on hepatic necrosis and total tract digestibility of energy, N, and amino acids have been well documented. However, the question remains if this is an effect on nutrient metabolizability alone or an effect on the intestine, or both, resulting in increased endogenous nutrient loss or reduced nutrient retention, or both. Therefore, a 2-wk feeding study with a crude AFLA culture was conducted with laying hens to measure endogenous losses and digestive functionality of the intestine. Hy-Line W36 hens were fed 1 of 4 experimental diets containing a crude AFLA culture from 20 to 22 wk of age. Diets were analyzed to contain 0, 0.6, 1.2, or 2.5 mg/kg of AFLA B(1). Dietary AFLA concentration had no effect on BW, egg production, or feed intake. Intestinal crypt depth (but not villus length) increased linearly with increasing AFLA concentration. Similarly, specific activity of the intestinal disaccharidase, maltase, increased quadratically by feeding up to 1.2 mg/kg of AFLA and declined at 2.5 mg/kg of AFLA (P <or= 0.022). Although there was no effect of AFLA on goblet cell number, density, or crude mucin excretion (P > 0.05), sialic acid excretion increased quadradically such that it was increased 12% when 0.6 and 1.2 mg/kg of AFLA was fed versus the control (P <or= 0.025). Digestibility of DM and N per hen per day were unaffected by AFLA. Feeding of 0.6 and 1.2 mg/kg reduced the apparent digestible and AME(n) of the hen by 10 and 4%, respectively (P <or= 0.025). Because intestinal characteristics (intestinal morphology, sialic acid production, and apparent digestible energy) were altered by AFLA feeding, it can be surmised that AFLA can have a direct or indirect effect, or both, on functionality of the gastrointestinal tract.