Endosymbiotic microflora of the vestimentiferan tubeworm ( Lamellibrachia sp.) from a bathyal cold seep.

Gutless vestimentiferan tubeworms are known to harbor endosymbiotic bacteria in a specialized tissue, the trophosome, which consists of lobules. The endosymbionts of vestimentiferans inhabiting sulfide-rich hydrothermal vents are monospecific for their host. In contrast, previous studies suggest that vestimentiferas of methane-rich seeps may host multispecific symbionts. Phylogenetic analysis and dot-blot hybridization of 16S ribosomal RNA genes (16S rDNA) detected 4 operational taxonomic units (OTUs) in the trophosome of the vestimentifera Lamellibrachia species from a bathyal methane-seep. The OTUs were closely related to 16S rDNA of the species belonging to alpha -Proteobacteria ( Sulfitobacter), beta- Proteobacteria ( Janthinobacterium), and gamma -Proteobacteria ( Acinetobacter and Pseudomonas). Localizations of the 4 OTUs within the trophosome were confirmed by in situ hybridization (ISH). ISH signals of the alpha-proteobacterial OTU were observed in the innermost zone of the trophosome lobules. In contrast, ISH signals of the beta- and gamma-proteobacterial OTUs were observed at the periphery of the lobules; however, whether they occur inside or outside the lobules remains unclear. These results support the possibility that the studied methane-seep tubeworm has a microflora composed of multispecific endosymbionts.