The aim of this study was to elucidate the effect of dietary fish oil (FO) and a blend of FO and hydrogenated palm oil (FOPO) on transport of fatty acids (FA) within plasma lipoproteins in lactating and non-lactating cows. Two trials were conducted (one with lactating and another with non-lactating dairy cows) in two 3 × 3 Latin squares that included three periods of 21 days. Dietary treatments for lactating cows consisted of a basal diet (Control; no fat supplement), and fat-supplemented diets containing FO (500 g/day/cow) and FOPO (250 FO + 250 g/day/cow hydrogenated palm oil). For non-lactating cows, dietary treatments consisted of a basal diet (Control; no fat supplement), and fat-supplemented diets containing FO (170 g/day/cow) and FOPO (85 FO + 85 hydrogenated palm oil g/day/cow). In lactating cows, compared with control and FOPO, FO increased C16:0, C18:3 cis-9, 12, 15, C18:2 cis-9, trans-11 and total saturated and polyunsaturated FA in plasma and increased C16:0, C18:2 cis-9, trans-11, total polyunsaturated and total polyunsaturated n-6 in high-density lipoprotein (HDL), whereas in non-lactating cows, compared with control and FOPO, FO increased C16:0, C18:1 trans-11, C18:2 trans-9, 12, C18:2 cis-9, trans-11, C20:5 n-3 and total saturated and polyunsaturated FA in plasma; C16:0, C18:1 trans-11, C18:1 cis-9, C18:2 trans-9, 12, C20:5 n-3 and total monounsaturated FA in HDL; and C18:1 trans-6-8, C18:1 trans-9, C18:1 trans-10, C18:1 trans-11, C18:3 cis-9, 12, 15 and C20:5 n-3 in low-density lipoprotein (LDL). FO increased C20:5 n-3 in plasma and lipoproteins in non-lactating cows and increased C18:3 cis-9, 12, 15 in plasma (in lactating cows) and LDL (in non-lactating cows). We concluded from results of this study that in bovine plasma, the LDL fraction appears to be the main lipoprotein transporting C18:1 trans isomers and is more responsive than other lipoprotein fractions to variation in supply of dietary lipids.