The macrofaunal community in the Haima cold seeps of the South China Sea was investigated, and trophic relationships were evaluated using carbon and nitrogen stable isotope signatures (δ13C and δ15N). A total of 30 macrofaunal species were identified in this study, most of which were collected from the Gigantidas haimaensis mussel beds. Only four macrofaunal species were collected from the Calyptogena marissinica clam beds, where the environment may not be suitable for the survival of most macrofauna. The bush of slim tubeworms (Sclerolinum sp.) might be an important nursery ground for mussels. G. haimaensis was the predominant species, and its average δ13C and δ15N values were −52.7‰ and 0.6‰, respectively. Compared with the stable isotope signatures of particulate organic matter (POM), it suggested that the food source of macrofauna in the Haima cold seeps was mainly originated from chemosynthetic autotrophs. In the mussel bed, most of the macrofauna had a large δ13C isotopic distance compared with G. haimaensis, suggesting that G. haimaensis was not a direct food source for most cold-seep macrofauna in the mussel bed. The variation in stable isotope signatures of macrofauna might be attributed to different proportions of methanotrophic and thiotrophic endosymbionts. Neolithodes brodiei and Bathynomus jamesi showed the highest δ13C and δ15N values. However, the large isotopic distance indicated that N. brodiei and B. jamesi might just occasionally visit the cold-seep ecosystem and their food sources were mainly from the upper ecosystem relying on photosynthesis. Overall, our results suggested that there was no significant predator-prey relationship among these macrofaunal species in the Haima cold seeps. This study provided important baseline data of stable isotope signatures for further studies to elucidate the trophic interaction among seep organisms.
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