The food sources of the main economic animals and trophic levels of biotic communities from an optimized culture model (Rhopilema–Sinonovacula constricta–Paralichthys olivaceus–Penaeus chinensis) in a sea water pond in Donggang, Liaoning province were studied using the stable carbon and nitrogen isotopes technique. The results indicated that the values of δ13C range from (−27.28 ± 0.35) ‰ to (−16.65 ± 0.20) ‰ and the values of δ15N range from (3.68 ± 0.23) ‰ to (13.91 ± 0.26) ‰, both of which exhibited significant fluctuations. The δ13C values of P. chinensis, Macrobrachium mipponensis and P. olivaceus were comparatively higher than those of other aquaculture animals, and the δ15N values of P. olivaceus was also comparatively higher. The contribution to the food sources of aquaculture animals (Rhopilema, S. constricta, P. olivaceus and P. chinensis) was analyzed by using the IsoSource software. The results indicated that fish meat had the greatest contribution to Rhopilema, S. constricta and P. chinensis; P. chinensis had the greatest contribution to P. olivaceus, followed by the M. mipponensis. The trophic level of the biotic communities under that optimized culture model in a sea water pond was 3.54, in which P. olivaceus was in the fourth level (3.54); P. chinensis, Synechogobius hasta, Ablennes anastomella, M. mipponensis, Ditrema temmincki Bleeker, Chelon haematocheilus were in the third trophic levels; Rhopilema, Engraulis japonius, S. constricta and zooplankton (1.00) in the second trophic levels, suspended matters (0.15) including phytoplankton, bacteria, humus, etc., were in the primary trophic levels.