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Context: Broiler chickens are exposed to various stressors, which can affect production performance. The use of dietary probiotics is one of the feasible methods to maintain performance, intestinal health, humeral immunity, and ameliorating physiological stress in the broiler. Aims: In all, 288 1-day-old male broiler chicks were used to evaluate the effect of probiotic supplementation on performance, morphology, and gene expression associated with immune responses in the starter phase of broiler chickens exposed to stress. Methods: In total, 288 1-day-old male broiler chicks (Ross 308) were assigned to evaluate the effects of Bacillus subtilis spore (BSS; Gallipro®200) supplementation at three levels (0, 0.8 × 105 colony-forming units (CFU) and 1.6 × 105 CFU/g feed) with corticosterone (CORT; 4 mg/kg BW at 7–9 days of age) or without CORT (oil) subcutaneous injection. The experiment was conducted in a completely randomised 2 × 3 factorial design. Key results: CORT injection and probiotic supplementation led to an increase and decrease respectively, in food conversion ratio compared with the control group (P < 0.05). CORT injections reduced the expression of TLR4, HSP70 and IgA in all parts of the intestine. However, the TLR4 gene expression increased in jejunum by CORT injections. BSS supplementation induced the expression of TLR4 in all sections of the intestine, both in stress and non-stress conditions. In stress-exposed chickens, IgA expression in the jejunum was affected by BSS supplementation. Interaction effects between CORT injections and BSS supplementation on the gene expression were significant in different parts of the intestine. Conclusions: Chicks exposed to stress conditions induced by CORT injection showed that the ileum was more susceptible than were the other parts, especially for TLR4 and HSP70 gene expression. BSS-supplemented birds showed more capability to face the stress condition, probably due to intestinal health and immunity-modulated effect of BSS. Implication: Formulating a ration with a probiotic can be more effective in stress conditions, through modulating immune-responsive genes in the intestine. The effects of various environmental stresses on the behavioral, physiological, and performance responses of birds have been studied for years. It has been shown that probiotics, also known direct-feed microbes, substantially influence nutrient digestibility, gut microflora and immunity, and inhibit pathogens and, ultimately, improve growth performance in broilers. Our findings suggested that formulating a ration with probiotic can be more effective in stress condition, with immune response genes modulating the effect in the intestine of broiler chicken. [ABSTRACT FROM AUTHOR]