Archaeal community structures in microhabitats in a deep-sea hydrothermal vent chimney structure were evaluated through the combined use of culture-independent molecular analyses and enrichment culture methods. A black smoker chimney was obtained from the PACMANUS site in the Manus Basin near Papua New Guinea, and subsamples were obtained from vertical and horizontal sections. The elemental composition of the chimney was analyzed in different subsamples by scanning electron microscopy and energy-dispersive X-ray spectroscopy, indicating that zinc and sulfur were major components while an increased amount of elemental oxygen in exterior materials represented the presence of oxidized materials on the outer surface of the chimney. Terminal restriction fragment length polymorphism analysis revealed that a shift in archaeal ribotype structure occurred in the chimney structure. Through sequencing of ribosomal DNA (rDNA) clones from archaeal rDNA clone libraries, it was demonstrated that the archaeal communities in the chimney structure consisted for the most part of hyperthermophilic members and extreme halophiles and that the distribution of such extremophiles in different microhabitats of the chimney varied. The results of the culture-dependent analysis supported in part the view that changes in archaeal community structures in these microhabitats are associated with the geochemical and physical dynamics in the black smoker chimney.Since the discovery of deep-sea hydrothermal vents in 1979 (12, 15), various microorganisms have been isolated from global deep-sea hydrothermal vent environments (26,27,29). Hyperthermophiles and thermophiles, including members of both the bacterial and archaeal domains, are the most frequently isolated microorganisms, and their physiological properties likely reflect the extraordinary environmental settings of the deep-sea hydrothermal vents (9,10,18,23,28,42,43,53,56,60). In addition, recent culture-independent molecular approaches have revealed the presence of as-yet-uncultivated thermophiles showing substantial phylogenetic diversity in the deep-sea hydrothermal vent environments (27,38,39,44,51). Relatively little is known about the ecological significance and geomicrobiological function of the extremophilic microbial communities found, as the occurrence, abundance, and distribution of the microbial communities associated with the formation of diverse geochemical and physical gradients remain to be elucidated.A deep-sea hydrothermal vent chimney, formed by chemical interaction between cold seawater and hot vent water, is a distinctive structure in the deep-sea hydrothermal fields and is largely composed of sulfide materials. In the chimney structures and the underlying sulfide mounds, steep environmental gradients of temperature, pH, oxidation-redox potential, and various chemicals can be formed by equilibration between the vent water and the seawater, and these provide diverse micro-