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Simple Summary  Different genotypes of slow- and fast-growing chickens have phenotypic changes in appearance, behavior, and productivity in response to artificial selection. Feed restriction and gut microbiota play a vital role in controlling food intake, nutrition, and health. However, little is known about how feed restriction, as a benefit or chronic stress, influences behavior, stress response, and gut microbial programming in slow- and fast-growing chickens. This study aimed to explore slow- and fast-growing chickens who had feed restricted to 70% of ad libitum or were given ad libitum feed for 30 days to evaluate the effects on behavior, stress response, and gut microbiota. We found that feed restriction can influence behaviors in both slow- and fast-growing breeds. Feed restriction to 70% for 30 days can influence stress response and gut microbiota composition, but some changes are evident only in slow- or only in fast-growing chickens. The study provides a better understanding of how artificial selection has affected chicken biology and their response to stress challenge.Abstract  This study aimed to explore the difference between two Chinese local broilers, one slow- and one fast-growing, in their response to a stress challenge. We conducted the study on slow- (Weining chicken) and fast-growing (Jinlinghua chicken) breeds, with 50 chickens from each breed either feed restricted to 70% for 30 days as a stress or given ad libitum to evaluate the effects on behavior, corticosterone, and microbial programming. Standing behavior was more frequent while exploration was less common in fast-growing breeds compared to slow-growing breeds. Food seeking and ingestion, exploration, and drinking increased, while resting decreased in the feed restricted treatments. There was no difference in corticosterone concentration between slow- and fast-growing chickens, but the level was affected by feeding treatments, and the interaction of breed and feed restriction. At the genus-level, the relative abundance of  Bacteroides  and  Lactobacillus  was higher, while  Cloacibacillus  and  Megasphaera  was lower in the slow-growing breed compared to the fast-growing breed. Feed restricted birds had a higher abundance of  Mucispirillum , but lower abundance of  Cloacibacillus ,  Clostridium XlVa  and  Clostridium IV . In conclusion, feed restriction to 70% for 30 days as a chronic stress stimulation caused more activity, elevated the stress response, and altered gut microbiota composition, but some changes were only evident in slow- or fast-growing chickens.

Feed Restriction Induced Changes in Behavior, Corticosterone, and Microbial Programming in Slow- and Fast-Growing Chicken Breeds