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Ecology of Sulfolobus acidocaldarius was studied in situ by the use of the immunofluorescence and immunodiffusion techniques. The fluorescent antibodies (FA) prepared against four strains of Sulfolobus were highly reactive against their homologous antigens. Two of the FA's were strain specific and the other two exhibited reciprocal corssreactions against each other's antigens, but immunodiffusion patterns showed that the two strains were not identical. The growth of a serologically distinct isolate in a hot spring was measured by immunofluorescence staining of immersion slides. On glass immersion slides Sulfolobus grew and formed colonies with a mean-doubling time of approximately 36 h. Immunofluorescence was applied to study the geographical distribution of two serologically different strains and to establish population composition of individual springs. One strain was found in all sites studied, and most springs contained more than one serologic type. Immunodiffusion was capable of detecting specific Sulfolobus antigens in hot springs which contained a high population of FA-reactive cells.

Summary: Sulfolobus attaches to sulphur deposited in acid hot springs by means of pili characterized as follows: (i) generally irregular shape with slight curves and bends; (ii) adhesiveness that enables bacteria to attach to sulphur; (iii) acid-stable, resistant to pH values as low as 2; (iv) heat-stable, resistant to temperatures as high as 75 °C. Sulfolobus attached to sulphur in nature and in culture eroded the sulphur crystal where the bacteria were attached. Sulfolobus undergoes the following two forms of attachment: (i) to sulphur by pili which separate the bacterium from the sulphur crystal and permit lateral movement of bacteria; (ii) to glass slides by firm adhesion of the wall to the surface of the slide. Attachment to sulphur in flowing springs enables Sulfolobus to colonize these low pH (2 to 3) high temperature (70 to 75 °C) habitats.

Sulfolobus is a new genus of bacteria characterized as follows: 1. generally spherical cells producing frequent lobes; 2. facultative autotrophy with growth on sulfur or on a variety of simple organic compounds; 3. unusual cell wall structure devoid of peptidoglycan; 4. acidophilic, pH optimum of 2–3 and range from 0.9–5.8; 5. thermophilic with temperature optimum of 70–75°C and range from 55–80°C (one strain grew at 85°C). The DNA base composition of five strains was determined by cesium chloride density gradient centrifugation and found to be 60–68% guanine plus cytosine. Sulfolobus apparently has no close relationship with any previously described bacteria, either heterotrophic or autrotrophic. Techniques are presented for distinguishing Sulfolobus from Thermoplasma, another genus of acidophilic thermophilic spherically shaped organisms. Sulfolobus has been isolated from a variety of natural acidic thermal habitats, both terrestrial and aquatic. Most isolations have been from habitats in Yellowstone National Park, but strains were also isolated from Italy, Dominica and El Salvador. It is suggested that Sulfolobus may be an important geochemical agent in the production of sulfuric acid from sulfur in high temperature hydrothermal systems.

Natural populations of Sulfolobus, a new genus of bacteria occurring in sulfur-rich, acid hot springs and soils, were found to oxidize large amounts of sulfur to sulfuric acid at temperatures up to 85 degrees C. These bacteria are important high-temperature geochemical agents in solfatara soils.