Seasonal migration for birds – distant migrants are the most energy intensive. Fat reserves accumulated in the bird’s body before migration and during migratory stopovers determine success of the long-distance flight. Lipids play a vital role both as a source of energy and as structural components of cell membranes. For most migrants to the speed and quality processes fat accumulation affects the feed ration in the field of migration stops. Fodder saturation with essential polyunsaturated fatty acids (PUFAs) is of great importance. Being paramount for physiological processes, these acids cannot be synthesized in the bird’s body. The proposed article is dedicated to the study of waders’ trophic relationships with their prey items, and the use of PUFAs as biochemical markers. This approach is based on the specificity of the fatty acids contained in the lipids of invertebrate to be used as food bird species studied. Significant amount NPZHK waders obtained from forage that can be considered PUFAs as biochemical markers to determine the range and diversity of food producing birds PUFAs ways, and also to study the food chain in ecosystems. A fatty acid spectrum (FAS) of the lipids common for nine littoral invertebrate species (Gammarus aequicauda, Idotea balthica, Artemia salina, Nerеis sp., Nerеis zonata, Theodoxces astrachanicus, Hydrobia acuta, Chironomus salinarius, Chironomus plumosus), which constitute the main component of waders’ diet at the migratory stopover sites in the Azov and Black Sea region, has been studied. Found that the largest amount of total lipids contained in Nereis zonata (4,6 %) and Artemia salina (4,4 %), the lowest amount of total lipids was observed in Chironomus (1,5–1,8 %), which implies that polychaete worms and Artemia salina, as a source of fat, are the most productive for waders. Our research has found that mollusks, polychaete worms, and Artemia salina are the most effective waders’ fodder in the PUFAs content. Mollusks contain the largest amount of PUFAs, their spectrum is ω3 and ω6 PUFAs, especially arаchidonic acid C20:4. Polychate worms are also characterized by a high PUFAs level; they serve for birds as a source of linolenic and linoleic acid groups. Artemia salina contains a large amount of eicosapentaenoic С20:5ω3 and docosahexaenoic С22:6ω3 acids, which getting to an organism of birds, participating in the formation of cell membranes, act as thermal stabilizer lipid bilayers, enhance stamina during long-distance flight. A high abundance of Artemia salina in the feeding areas permit tundra waders to use them as a prey item, which can fulfill the bird’s body with a required amount of fatty acids in a short time. We have established an influence of some environmental factors, as water temperature and salinity, on the lipids FAS of littoral invertebrates. Spectrum analysis of polyene fatty acids in the lipids closely related species of invertebrates living in different salinity water showed that the content of PUFAs in the lipid depend on their food spectrum, and the environmental conditions. Therefore, anthropogenic pressure and changes of water hydrological regime may affect PUFAs content in the lipids of littoral invertebrates. In its turn, this factor may change alimentary behavior and migratory strategy of the birds, which use migratory stopover sites in the region in question. Shallow waters of the Azov-Black Sea region are characterized by different climatic characteristics and a large reserve of phytoplankton. This explains the mass character species studied of invertebrates to feeding areas and their use as basic prey items, many species of waders.