The biochemical composition and biometry of different Artemia salina stages were determined in four ponds of increasing salinity (M1, M2, M3 and B1) in the Sfax solar saltern (Tunisia). Results showed the dominance of saturated fatty acids, which made up 48 to 57% of total fatty acids (FAs). Polyunsaturated fatty acids (PUFAs) 22:6(n-3) docosahexaenoic acid (DHA) and 20:5(n-3) eicosapentaenoic acid (EPA) represented on average only 3.1 and 4.0% of total FAs respectively. A. salina nauplii, cysts and metanauplii, in ponds M1, M2 and B1 respectively, were found to have optimal DHA/EPA ratios (>2) for use as live feed for invertebrate and fish larvae. Significant inter-pond variation in DHA/EPA levels was also recorded. The predominant FAs in Artemia were negatively correlated with both temperature and salinity. FA and protein contents were strongly affected by high temperatures (>30 ◦C) and probably by food sources (e.g., Dunaliella salina) (r = 0.9, n = 27). The density of Artemia salina was positively correlated with protein content in pond B1. The high DHA/EPA ratios (1 to 3.3) found in this study indicate that Sfax Artemia could be a valuable food source for larvae in large marine hatcheries and also for some aquarium species.