Abstract Near (NIR) and medium (MIR) infrared reflectance spectroscopy (IR) predictions of fatty acid (FA) composition, expressed as g/kg of milk or g/100g of FA, on fresh and thawed milk were compared. Two-hundred-and-fifty bulk cow milks, collected from 70 farms in northwest Italy, were scanned by MIR in liquid form and by NIR in liquid and oven-dried forms. MIR and NIR FA (g/100g FA) predictions on oven-dried milk were similar for the sum of even chain-saturated FA (ECSFA), odd chain-FA (OCFA), unsaturated FA (UFA), conjugated linoleic acid (CLA), n-3 FA, and C18:1cis9 to C16 ratio. The monounsaturated FA (MUFA), n-6 to n-3 ratio, polyunsaturated FA (PUFA), and n-6 FA were predicted better by NIR on oven-dried milk. The NIR showed worse predictions than MIR for almost all FA, when expressed as g/kg of milk. The NIR predictions on fresh liquid and oven-dried milk were similar, but the reliability decreased for thawed liquid milk. The high performance shown by NIR and MIR allows their use for routine milk FA composition recording.