Since sheep's milk is mainly used for cheese making and milk chemical composition and fatty acids (FA) profile affect cheese yield and quality, the objective of this study was to determine the effects of different feeding levels on milk chemical composition and FA profile, as well as on plasma FA profile, and on insulin and leptin concentrations. Twenty-four sheep were assigned to three homogeneous sub-groups. Throughout the experimental period each group was fed the same diet but in quantities which met 70% (under-feeding), 100% (control) and 130% (over-feeding) of their respective energy and crude protein requirements. The results showed that the underfed sheep had higher milk fat content compared with overfed. In blood plasma the concentrations of C18:0 and C18:1 in the underfed sheep were significantly higher compared with control and overfed sheep. The concentrations of leptin and insulin were significantly higher in overfed compared with underfed sheep. Underfeeding reduced the concentrations of short chain FA (SCFA) and medium chain FA (MCFA) and increased that of C18:0 and mono unsaturated FA (MUFA) in sheep milk fat compared with controls and overfed. The concentrations of C18:0, long chain FA (LCFA) and monounsaturated FA (MUFA) in milk were significantly higher and those of SCFA, MCFA and saturated FA (SFA) significantly lower in the underfed compared with the overfed sheep. In conclusion, long term under- and over-feeding affected the sheep milk chemical composition and FA profile which consequently has an impact on milk products yield (cheese and yogurt) and quality (human health).