Immunodiffusion analysis of membranes of Thermoplasma acidophilum.
Isolates of Thermoplasma acidophilum were divided into serological groups based on immunodiffusion studies of solubilized-membrane preparations.
Biomedical subjects
Publications and source records attributed to T D Brock.
Isolates of Thermoplasma acidophilum were divided into serological groups based on immunodiffusion studies of solubilized-membrane preparations.
A technique was developed for measuring (14)CO(2) uptake by chemolithotrophic bacteria directly in pyritic materials associated with coal and coal refuse. There was good correlation between (14)CO(2) uptake, as determined by this technique, and the most probable number of iron-oxidizing bacteria. Maximal (14)CO(2) uptake occurred in coal refuse material 2 to 3 years old, and only slight incorporation was demonstrated in fresh material or material 40 years old. Samples taken from the surface of the coal refuse pile always demonstrated maximal (14)CO(2) uptake, and in most samples, only slight activity was demonstrated at depths below 8 to 10 cm. Optimal uptake of (14)CO(2) by natural samples occurred at 20 to 30 C and at a moisture content of between 23 and 35%. In addition to chemolithotrophic bacteria, heterotrophic fungi and yeasts were also routinely isolated in high numbers from acidic coal refuse. In contrast, acidophilic, heterotrophic bacteria were either absent or present in low numbers in such acidic samples.
Turnover times for water passing through several Sulfolobus acidocaldarius-containing springs were determined by measuring the dilution rates of small amounts of sodium chloride that were added to the springs. Chloride was diluted out exponentially, while concentrations of the bacteria remained constant. Additionally, temperature, pH, and chemical composition of the springs also remained constant during the time that the chloride was being diluted. The springs are thus steady-state systems, and since the rates of bacterial growth must be at least equal to the chloride dilution rates, minimal doubling times for the bacterial populations can be calculated. Half-times for chloride dilution, equivalent to bacterial doubling times, were on the order of 10 to 20 h for springs ranging in volume from about 20 to 2,000 liters, but approximately 30 days for two larger springs of about 1 million liters. Formaldehyde-fixed cells of a serologically distinguishable strain of S. acidocaldarius were also added as markers to four of the smaller springs, and the dilution rates of these bacteria were compared with the chloride dilution rates. The rates agreed reasonably well, thus verifying the growth rates obtained from the chloride dilution rates. In three springs, exponential growth was studied by draining the springs and allowing them to refill with bacteria-free water. Exponential doubling times were on the order of a few hours, much more rapid than steady-state doubling times. The methods used in this work may have wider utility in aquatic environments.
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Observations on a wide variety of acidic environments, both natural and man-made, reveal that blue-green algae (Cyanophyta) are completely absent from habitats in which the pH is less than 4 or 5, whereas eukaryotic algae flourish. By using enrichment cultures with inocula from habitats of various pH values, the absence of blue-green algae at low pH was confirmed.
Thermophilic bacteria resembling Thermus aquaticus were isolated from hot water taken from domestic and commercial hot-water tanks. Cold water from the same locations never yielded thermophilic bacteria, suggesting that the bacteria were growing in the tanks. In contrast to the T. aquaticus isolates from hot springs, the present isolates were rarely pigmented. In general, the hotter sources more frequently yielded bacteria.
Oxidation of elemental sulfur by Sulfolobus acidocaldarius, an autotroph which grows at high temperatures and low pH, was examined by use of (35)S-labeled elemental sulfur. When cultured at pH 3.2 and 70 C, S. acidocaldarius oxidized elemental sulfur essentially quantitatively to sulfuric acid. Oxidation rate paralleled growth rate and decrease in pH of the culture medium. Elemental sulfur was not oxidized under these conditions if the culture was poisoned with formaldehyde. During the growth phase, the proportion of cells attached to the sulfur crystals increased progressively, and in the later phases of growth over 10 times more cells were attached to sulfur than were free. Doubling times for eight strains growing on elemental sulfur varied from 37 to 55 h. The organism grows much more rapidly on yeast extract than on sulfur. In a medium containing both sulfur and yeast extract, sulfur oxidation was partially inhibited, although growth was excellent.
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An upper temperature limit near 60 degrees for eukaryotic organisms is documented by results of a systematic search for fungi able to grow at higher temperatures. Samples from hot springs, thermal soils, self-heating coal waste piles, and other natural and man-made heated habitats did not yield fungi when enrichments were done at 62 degrees , whereas fungi able to grow at 55-60 degrees can be readily isolated from such habitats. Earlier work had shown that eukaryotic algae are also absent from environments with temperatures above 55-60 degrees . It is suggested that the failure of eukaryotes to evolve members able to grow at higher temperatures is due to their inability to form organellar membranes that are both thermostable and functional.
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Uptake of (14)CO(2) in soils due to algae or sulfur-oxidizing bacteria was examined by incubation of soil samples with gaseous (14)CO(2) and subsequent chemical oxidation of biologically fixed radioactive isotope to (14)CO(2) for detection with a liquid scintillation counting system. The (14)CO(2) was added to the soil in the gas phase so that no alteration of the moisture or ionic strength of the soil occurred. Wet oxidation of radioactive organic matter was carried out in sealed ampoules, and the (14)CO(2) produced was transferred to a phenethylamine-liquid scintillation counting system with a simply constructed apparatus. The technique is inexpensive and efficient and does not require elaborate traps since several possible interfering factors were found to have no harmful effects. Experiments in coal mine regions and in geothermal habitats have demonstrated the ecological applicability of this technique for measurement of CO(2) fixation by sulfur-oxidizing bacteria and soil algae.
Hot acid soils in Yellowstone National Park are rich in elemental sulfur and harbor extensive populations of sulfur-oxidizing bacteria. Thiobacillus thiooxidans is found at temperatures below 55 C, and at temperatures from 55 to 85 C Sulfolobus acidocaldarius is present. The distribution of these bacteria as a function of temperature was measured by a most-probable-number dilution method, and their activity in situ was assessed by use of a new technique permitting measurement of (14)CO(2) fixation. From these data it is concluded that sulfur-oxidizing bacteria are responsible for production of sulfuric acid in these acidic thermal habitats. Physical and chemical parameters of this unusual soil habitat were also measured and are described.
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Thermus aquaticus contains four major fatty acids, iso-C(15) (28%), iso-C(16) (9%), normal-C(16) (13%), and iso-C(17) (48%), when grown at 70 C, as determined by gas chromatography and mass spectrometry. Small amounts of iso-C(12), normal-C(12:1), iso-C(13), normal-C(14), iso-C(14), and normal-C(15:1) were also detected. A change in growth temperature (50 to 75 C at 5-C intervals) affects a shift in the proportions of some of the fatty acids. The proportions of the monoenoic and branched-C(17) fatty acids decreased and the proportions of the higher-melting iso-C(16) and normal-C(16) fatty acids increased. Cells grown at 75 C contained 70% more total fatty acids than cells grown at 50 C. The largest increases, in absolute amounts, were in the content of iso-C(16) and normal-C(16) fatty acids, with only a 1.6-fold increase in the major iso-C(15) and iso-C(17) fatty acids. There was a 2.5-fold decrease in normal-C(15:1) and at least a 24-fold decrease in anteiso-C(17), which is present at 50 and 55 C but not at higher temperatures. There was no difference in proportion or amount of fatty acids between exponential and stationary-phase cells grown at 70 C. When cells were grown on glutamate instead of yeast-extract and tryptone at 70 C, the total fatty acid content remained constant, but there was an increase in the proportions of iso-C(16) and normal-C(16) fatty acids concomitant with a decrease in the proportions of the iso-C(15) and iso-C(17) fatty acids.