A Combined Immunofluorescence-DNA-Fluorescence Staining Technique for Enumeration of Thiobacillus ferrooxidans in a Population of Acidophilic Bacteria

Abstract

An antiserum raised against whole cells of Thiobacillus ferrooxidans was allowed to react with a variety of acidophilic and nonacidophilic bacteria in an enzyme-linked immunosorbent assay and an indirect immunofluorescence assay. Both experiments demonstrated that the antiserum was specific at the species level. This preparation was used to evaluate the role of T. ferrooxidans in the microbial desulfurization process. Leaching experiments were performed, and the numbers of T. ferrooxidans cells and other bacteria were estimated by using a combined immunofluorescence-DNA-fluorescence staining technique that was adapted for this purpose. Nonsterile coal samples inoculated with T. ferrooxidans yielded high concentrations of soluble iron after 16 days. After this period, however, T. ferrooxidans cells could no longer be detected by the immunofluorescence assay, whereas the DNA-fluorescence staining procedure demonstrated a large number of microorganisms on the coal particles. These results indicate that T. ferrooxidans is removed by competition with different acidophilic microorganisms that were originally present on the coal.

Evaluation of Leptospirillum ferrooxidans for Leaching

“…In the former case, the results are incomplete because of the absence of the effect of L. ferrooxidans whereas in the latter case L. ferrooxidans may have inadvertently contributed to the leaching. Muyzer et al (21) reported such a finding when an immunofluorescence-DNA-fluorescence staining technique for monitoring a leaching experiment on desulfurization of coal was used. At 16 days after the inoculation of nonsterile coal samples, the autochthonous flora had overgrown the inoculated T. ferrooxidans and effectively removed pyrite.…”

Competition between heterotrophic and autotrophic nitrifiers for ammonia in chemostat cultures

“…As 7". pantotropha has a much lower K~ for oxygcn (1-2/x M [23]) than N. europaea (15)(16)(17)(18)(19)(20) /.tM [24]), it would appear that compctition bctween the two bacteria for oxygen might become a significant factor. This would imply that the hetcrotroph should have a competitive advantage at low dissolved oxygen concentrations.…”

Floating Filters, a Novel Technique for Isolation and Enumeration of Fastidious, Acidophilic, Iron-Oxidizing, Autotrophic Bacteria

“…All liquids used in the fluorescence technique were filtered through a 0.2 ,um filter prior to use. Immunofluorescence staining with specific antiserum against T. ferrooxidans was carried out as described by Muyzer et al (8).…”