This study aimed to investigate the effects of early or late feeding strategies and prebiotic, on immune responses and gut health during the early life stage of broiler chickens. A total of 240 day-old male broiler chicks were used in a 2 × 3 factorial arrangement of treatments that comprised 2 feeding strategies (early or late) and 3 levels of prebiotic (0, recommended dosage or three times the recommended dosage) in a completely randomized design with 4 pen replicates and 10 broilers per each. Compared to broiler chickens that had early access to feed, delayed access to feed resulted in an increased population of Escherichia coli and a decreased population of Lactobacillus spp. and Bifidobacterium spp. in the ileum (P < 0.05). Additionally, delayed access to feed led to a decrease in villus height, crypt depth, villus height: villus width ratio, goblet cell density, and mucin 2 gene expression in the ileum (P < 0.05). The supplementation of prebiotics in both the late and early feeding strategy groups resulted in increased villus height, crypt depth, goblet cell density, mucin 2 gene expression, and antibodies against Infectious Bursal Disease (IBD). Additionally, it led to an improvement in the foot web thickness index (P < 0.05). Furthermore, it resulted in a significant decrease in the population of Escherichia coli, while the populations of Lactobacillus spp. and Bifidobacterium spp. in the ileum were significantly increased (P < 0.05). Therefore, this study suggests that incorporating prebiotics in the starter diet can effectively enhance immune responses and promote gut health, regardless of the feeding strategy (early or late). In conclusion, this study demonstrates the potential benefits of incorporating prebiotics into poultry diets to alleviate the detrimental effects of delayed access to feed and improve gut health during the early life stage of broiler chickens.