During the 2010/2011 Exploration vessel Nautilus expedition to the Mediterranean Sea, samples of Lamellibrachia (Siboglinidae, Annelida) were imaged in situ and collected from hydrothermal vent and methane "cold seeps." An analysis of these Lamellibrachia and their endosymbiotic thioautotrophic gammaproteobacteria reveals two distinct endosymbiont phylotypes. Phylotype 1 was present in Lamellibrachia specimens from 947 m at the Eratosthenes seamount seep (a seep off Cyprus in the Eastern Mediterranean), and Phylotype 2 was found in siboglinids from 618 m at a hydrothermal vent within Palinuro volcanic complex in Tyrrhenian Sea. Both phylotypes coexist in siboglinids at 1,036 m from the Palmachim disturbance, a cold seep in the Eastern Mediterranean's Levantine basin. Our results, combined with existing knowledge of siboglinid host and endosymbiotic bacteria biogeography, reveal that two major groups of endosymbionts coexist within lamellibranchids and escarpids. The phylogenetic clustering of these bacteria is primarily influenced by geographic location, rather than selection by the siboglinid host.