Fatty acids, vitamins and minerals in milk are important for the human consumer, the calf and the cow. Studies indicate that milk from organic and conventional dairy herds may differ in these aspects. The aim of this study was therefore to investigate whether there are differences in the fatty acid composition and concentration of vitamins and selenium in milk between organic and conventional herds in Sweden. Bulk tank milk was sampled in 18 organic and 19 conventional dairy herds on three occasions during the indoor season 2005–2006. Herd characteristics were collected by questionnaires and from the official milk recording scheme. Multivariable linear mixed models were used to evaluate the associations between milk composition and type of herd, while adjusting for potential confounders and the repeated observations within herd. In addition to management type, variables included in the initial models were housing type, milk fat content, herd size, average milk yield and time on pasture during summer. The median concentration of conjugated linoleic fatty acids (CLA) was 0·63% in organic compared with 0·48% in conventional herds, the content of total n-3 fatty acids was 1·44% and 1·04% in organic and conventional milk, respectively, and the content of total n-6 fatty acids was 2·72% and 2·20% in organic and conventional milk, respectively. The multivariable regression models indicated significantly higher concentrations of CLA, total n-3 and n-6 fatty acids in organic milk and a more desirable ratio of n-6 to n-3 fatty acids, for the human consumer, in organic milk. The multivariable models did not demonstrate any differences in retinol, α-tocopherol, β-carotene or selenium concentrations between systems. Median concentrations of α-tocopherol were 0·80 μg/ml in organic and 0·88 μg/ml in conventional milk, while for β-carotene the median concentrations were 0·19 and 0·18 μg/ml, respectively; for retinol, the median concentration was 0·32 μg/ml in both groups; the median concentrations of selenium were 13·0 and 13·5 μg/kg, respectively, for organic and conventional systems.