The gastrointestinal tract plays a crucial role in nutrient absorption, secretion, and motility, ensuring proper digestion and overall homeostasis. Regulation of this complex system involves the coordination of various communication pathways, including neural and humoral mechanisms. One such mechanism is the endocannabinoid system (ECS), a signalling network comprising endogenous cannabinoids, receptors, and enzymes involved in the regulation of physiological processes in mammals and non-mammalian species. While extensive research has been conducted on the ECS in monogastric animals, limited information is available on its presence and distribution in cattle. This study aimed to investigate the distribution and localization patterns of cannabinoid receptors type 1 (CB1R) and type 2 (CB2R) and transient receptor potential vanilloid type 1 (TRPV1) in the bovine small intestine. The study included immunohistochemical analysis of intestinal tissue samples from Polish Holstein-Friesian breed bulls. Gene expression levels of