Changes in milk fat composition influence its nutritional quality as well as the technological properties of butter. The impact of feed on fat composition is well known; however, limited information is available on the genetic variability of fatty acids in bovine milk. The overall aim of this PhD thesis was to study the genetic variability of fatty acid profile in bovine milk and fat. This type of research needs a large amount of data. Expensive reference analysis is used to measure the fatty acid contents in fat. The first objective of this thesis was to develop an alternative method that could be faster and cheaper than traditional methods. Calibration equations predicting the contents of fatty acid from mid-infrared spectrum were established. The contents of saturated and monounsaturated fatty acids, omega-9, fatty acids with short, medium and long chain were the best predicted. Thanks to the implementation of this method in the Walloon routine milk recording, more than 20,000 milk samples were analyzed. This database permitted to model the variation of fatty acid contents in milk and fat. From these models, the genetic variability of fatty acid profile was shown. The complexity of models increased throughout this project due to the increase of new available data. Differences across 7 dairy breeds were estimated using single and multi-trait mixed models. Milk fat and delta-9 desaturase activity of Jersey and dual purpose Belgian Blue differed significantly from Holsteins. Therefore, the choice of a given breed could modify the fat composition. Heritability values obtained for studied fatty acids with multi-trait mixed model ranged from 0.05 to 0.42. Higher values were observed for saturated compared to unsaturated fatty acids. Moderate heritability estimates were observed for the activity of delta-9 desaturase (0.20) and the hardness of butter (0.27). These two traits were estimated by specific fatty acid ratios. The heritability observed using a multi-trait random regressions test day model for the content of saturated fatty acids (0.42) was similar to the one observed for the percentage of fat (0.37). Considering the impact of selection on fat content, the selection could have a great impact on fat composition. High genetic correlations were observed between some fatty acids having similarities in their synthesis. Heritability and correlations varied through the duration of the lactation. Due to the large number of fatty acids, the estimation of an index, which includes the proper fatty acid profile, could be interesting for a future selection program. This PhD thesis provides the background required by future studies to estimate the impact of animal selection on milk fat composition.