Fatty acids in fish can arise from two sources: synthesis de novo from non-lipid carbon sources within the animal, or directly from dietary lipid. Acetyl-CoA derived mainly from protein can be converted to saturated fatty acids via the combined action of acetyl-CoA carboxylase and fatty acid synthetase. The actual rate of fatty acid synthesis de novo is inversely related to the level of lipid in the diet. Freshwater fish can desaturate endogenously-synthesized fatty acids to monounsaturated fatty acids via a delta 9 desaturase but lack the necessary enzymes for complete de novo synthesis of polyunsaturated fatty acids which must therefore be obtained preformed from the diet. Most freshwater fish species can desaturate and elongate 18:2(n-6) and 18:3(n-3) to their C20 and C22 homologues but the pathways involved remain ill-defined. Cyclooxygenase and lipoxygenase enzymes can convert C20 polyunsaturated fatty acids to a variety of eicosanoid products. The dietary ratio of (n-3) to (n-6) polyunsaturated fatty acids influences the pattern of eicosanoids formed. The beta-oxidation of fatty acids can occur in both mitochondria and peroxisomes but mitochondrial beta-oxidation is quantitatively more important and can utilise a wide range of fatty acid substrates.