Fatty acid composition, conjugated linoleic acid and cholesterol contents in the muscles of three freshwater fish species (Barbus plebejus escherichi, Capoeta capoeta capoeta and Rutilus rutilus) were determined under natural extreme temperate (July) and cold (January) conditions. The aim of the study was to determine whether there were differences in these components of the muscle lipids among these three fish species under extreme natural conditions. Samples were analyzed using gas chromatography. Palmitic, oleic, docosahexaenoic and eicosapentaenoic acids were the predominant fatty acids in all fish in both months. The percentages of polyunsaturated fatty acids, n - 3 polyunsaturated fatty acids, n - 6 polyunsaturated fatty acids and eicosapentaenoic + docosahexaenoic acids in the muscle of B. plebejus escherichi and C. capoeta capoeta were significantly higher in January (P < 0.05) than in July. The ratio of n - 6 to n - 3 polyunsaturated fatty acids was lower than 0.60 in all fish species, with C. capoeta capoeta showing the lowest ratio in January (0.36). The levels of cholesterol and conjugated linoleic acid ranged from 103.46 to 150.10 mg/100 g oil and from 16.27 to 35.45 mg/100 g oil, respectively, for all samples in both months. There were no statistical differences in cholesterol levels among the three fish species in July and January. Conjugated linoleic acid contents were significantly higher in January in B. plebejus escherichi and C. capoeta capoeta. Of the three species tested, the extreme temperate and cold conditions affected B. plebejus escherichi the most.