Olive pomace (OP) is a residue of the oil extraction of olive fruit, which can be used in animal feed as one of the cheap and nutritious residues. This study was conducted to investigate the effect of olive pomace in three levels of OP0, OP7.5 and OP15% on milk and yogurt composition and milk fatty acid profile of dairy buffalo. Milk fat and SNF (solids not fat) increased in olive cake treatments (p < 0.05) but milk production, protein, and pH were not different (p > 0.05). The acidity, microbial load, Staphylococci, and SNF of yogurt produced from buffalo’s milk did not influence by treatments (p > 0.05). Yogurt fat decreased (p < 0.05) in OP7.5 and increased (p < 0.05) in OP15 buffalos. Protein concentration in yogurt dropped by olive pomace diets, and coliforms were the highest for OP15 olive pomace (p < 0.05). The concentration of short and medium chains fatty acids (C4:0, C8:0, C10:0, C12:0), linolenic acid, and conjugated fatty acids (CLA isomers) of buffalo’s milk in OP15 is more than other treatments (p < 0.05). The replacement of olive pomace with wheat flour increased milk production of dairy buffaloes. The concentration of short and medium-chain fatty acids and un-saturated and conjugated fatty acids increased in OP15 buffalos. Furthermore, a lower saturated/unsaturated ratio and atherogenic index suggest an improvement in nutritional characteristics of milk buffaloes. Therefore, it’s recommended in the diet of dairy buffaloes.