Fasting older broiler chickens (>7 d of age) enlarges the intestinal tight junction (TJ) pore size, resulting in high paracellular intestinal permeability. Broiler chickens often do not receive feed and water (nutrition) directly after hatch, which may result in fasting up to 72 h of age. Whether perinatal fasting affects intestinal permeability is minimally studied. We therefore investigated whether delayed access to nutrition after hatch increases intestinal permeability, compared with broilers receiving early access to nutrition. Therefore, 432 hatched broilers received nutrition 72 h after hatch (delayed nutrition [DN]) or directly after hatch (early nutrition [EN]) and were reared under similar conditions until 14 d of age. Two hours after application of an oral pulse dose (3.85 mg) of fluorescein isothiocyanate-dextran (4000 Da) at 4, 10, and 14 d of age, blood plasma concentrations of the marker were measured in 24 to 36 broilers per treatment and time point. Marker concentration in plasma did not differ between DN and EN broilers at any age. The villus width measured in at least 8 broilers per treatment was smaller in DN than in EN broilers at 4 d for both the ileum (92 ± 3 μm vs. 121 ± 4; P < 0.001) and colon (100 ± 3 vs. 120 ± 4; P < 0.01). Real-time quantitative PCR revealed that the expression of TJ protein claudin 3 in the ceca was elevated in DN, compared with EN broilers at 4 d of age, whereas that of zonula occludens 1 in the ileum was reduced. Expression of host defense-related genes was reduced in DN, compared with EN broilers, in the ileum (cyclo-oxygenase 2, mucin 2) and ceca (interleukin 1β, cyclo-oxygenase 2). We conclude that 72-hour DN reduced the BW up to 14 d of age, coinciding with transient effects on the villus width in the ileum and colon, and divergent expression of genes involved in TJ formation and host defense. These effects likely reflect the delayed onset of intestinal and immune development in DN, compared with EN broilers, while DN does not fundamentally alter intestinal permeability.