Buffalo meat is a nutritious alternative to beef, offering the advantages of lower fat and cholesterol levels. This study compared the fat characteristics of five crossbred water buffalo and five crossbred cattle. The ten growing animals averaged 22 months of age and were fed a diet of 50% Napier grass and 50% commercial concentrate, administered in two equal portions at 7:00 and 16:00 hours. During a 16-week fattening period, the total plasma cholesterol concentrations in water buffalo were significantly lower (P < 0.05) than in cattle. Additionally, the ether extract and cholesterol content in muscle tissue were significantly lower in water buffalo (P < 0.05). However, there were no significant differences between the species in relative leptin mRNA levels in fat tissue (P > 0.05) or adipocyte diameter (P > 0.05). In terms of fatty acid composition, monounsaturated fatty acid levels were similar (P > 0.05), but water buffalo exhibited higher levels of polyunsaturated fatty acids than cattle (P < 0.05). Overall, buffalo meat presents lower fat, cholesterol content, and higher essential fatty acids than beef, making it a potentially healthier choice for health-conscious consumers.