Unnatural diet composition and frequent feeding regimes may play an aetiological role in the multiple diseases prevalent in captive cheetahs. This study investigated the responses of captive-born (hand-reared) cheetahs (n = 6) to a reduced feeding frequency schedule. During the 3-week treatment period, the study cheetahs were fed four once-daily meals per week and two daily rations six days a week, subsequently, during the control period of equal duration. Total weekly food intake was maintained throughout the study. Variations in body temperature (Tb), heart rate (HR), locomotor activity (LA), behaviour, faecal glucocorticoid metabolite concentration, and faecal consistency score (FCS) were measured. Less frequent feeding resulted in higher FCS (p < 0.01) and LA (p < 0.0001) and lower HR (p < 0.0001) among the studied cheetahs. The study found that their HR was higher when the cheetahs were fed (p < 0.0001) than on days when they were not. Additionally, Tb (p < 0.0001) and HR (p < 0.05) were higher during feeding, suggesting a psychological excitation to food consumption. The results of the FCS analysis indicate that the more natural feeding pattern could have benefited the studied cheetahs’ GI health without a significant behavioural or physiological stress response to the change in feeding frequency.