Culturable thermophilic microorganisms were enriched from samples collected from Lau Basin hydrothermal vents in artificial seawater medium at 45 °C and pH 7.0. Microbial diversities of the enriched communities were defined by performing a restriction fragment length polymorphism (RFLP) analysis of 16S rRNA gene sequences with enzymes MspI and Hin6 I. A total of 14 phylotypes have been detected by the RFLP patterns identified for 16S rRNA clone libraries of the enrichment. Analysis of sequences showed that at least four bacterial divisions presented in the clones libraries. The phyla Proteobacteria and Firmicutes were the most dominant groups. The majority of the sequences included in this analysis affiliated with Gamma Proteobacteria (71%) and Bacillus (23%). Scanning electron micrographs revealed that there were abundant rod and coccoidal forms encased in sulphur and sodium chloride precipitate. These results revealed that there were a diversity of moderate thermophilic bacterial populations thrived in Lau Basin hydrothermal vents that were previously not detected by either molecular retrieval or strain purification techniques.