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. 1982 Feb;43(2):338–344. doi: 10.1128/aem.43.2.338-344.1982

Determination of the Number of Respiring Thiobacillus ferrooxidans Cells in Water Samples by Using Combined Fluorescent Antibody-2-(p-Iodophenyl)-3-(p-Nitrophenyl)-5-Phenyltetrazolium Chloride Staining

Katherine H Baker 1,, Aaron L Mills 1
PMCID: PMC241828  PMID: 16345938

Abstract

Fluorescent antibody staining was combined with 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride reduction in a procedure termed FAINT to allow for the direct microscopic determination of specific actively respiring populations of bacteria in a variety of aquatic habitats. The FAINT procedure is simple, precise, and appropriate for use in a wide variety of autecological studies. The distribution of Thiobacillus ferrooxidans was examined by FAINT enumerations in both acidic and nonacidic sites. Comparisons among the FAINT technique and fluorescent antibody staining alone or most-probable-number determinations in 9K broth showed that the use of most-probable-number determinations resulted in an underestimation of the number of viable cells by one to three orders of magnitude, whereas fluorescent antibody counts resulted in an overestimation of the number of viable T. ferrooxidans. The amount of difference was not consistent but varied, depending on the sample site.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Apel W. A., Dugan P. R., Filppi J. A., Rheins M. S. Detection of Thiobacillus ferrooxidans in acid mine environments by indirect fluorescent antibody staining. Appl Environ Microbiol. 1976 Jul;32(1):159–165. doi: 10.1128/aem.32.1.159-165.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bohlool B. B., Brock T. D. Immunofluorescence approach to the study of the ecology of Thermoplasma acidophilum in coal refuse material. Appl Microbiol. 1974 Jul;28(1):11–16. doi: 10.1128/am.28.1.11-16.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bohlool B. B., Schmidt E. L. Nonspecific staining: its control in immunofluorescence examination of soil. Science. 1968 Nov 29;162(3857):1012–1014. doi: 10.1126/science.162.3857.1012. [DOI] [PubMed] [Google Scholar]
  4. Casida L. E., Jr Microbial metabolic activity in soil as measured by dehydrogenase determinations. Appl Environ Microbiol. 1977 Dec;34(6):630–636. doi: 10.1128/aem.34.6.630-636.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cherry W. B., Pittman B., Harris P. P., Hebert G. A., Thomason B. M., Thacker L., Weaver R. E. Detection of Legionnaires disease bacteria by direct immunofluorescent staining. J Clin Microbiol. 1978 Sep;8(3):329–338. doi: 10.1128/jcm.8.3.329-338.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fliermans C. B., Bohlool B. B., Schmidt E. L. Autecological study of the chemoautotroph Nitrobacter by immunofluorescence. Appl Microbiol. 1974 Jan;27(1):124–129. doi: 10.1128/am.27.1.124-129.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fliermans C. B., Cherry W. B., Orrison L. H., Thacker L. Isolation of Legionella pneumophila from nonepidemic-related aquatic habitats. Appl Environ Microbiol. 1979 Jun;37(6):1239–1242. doi: 10.1128/aem.37.6.1239-1242.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fliermans C. B., Schmidt E. L. Autoradiography and immunofluorescence combined for autecological study of single cell activity with Nitrobacter as a model system. Appl Microbiol. 1975 Oct;30(4):676–684. doi: 10.1128/am.30.4.676-684.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Francisco D. E., Mah R. A., Rabin A. C. Acridine orange-epifluorescence technique for counting bacteria in natural waters. Trans Am Microsc Soc. 1973 Jul;92(3):416–421. [PubMed] [Google Scholar]
  10. Harvey R. W., Young L. Y. Enumeration of particle-bound and unattached respiring bacteria in the salt marsh environment. Appl Environ Microbiol. 1980 Jul;40(1):156–160. doi: 10.1128/aem.40.1.156-160.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hobbie J. E., Daley R. J., Jasper S. Use of nuclepore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol. 1977 May;33(5):1225–1228. doi: 10.1128/aem.33.5.1225-1228.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jones J. G., Simon B. M. An investigation of errors in direct counts of aquatic bacteria by epifluorescence microscopy, with reference to a new method for dyeing membrane filters. J Appl Bacteriol. 1975 Dec;39(3):317–329. doi: 10.1111/j.1365-2672.1975.tb00578.x. [DOI] [PubMed] [Google Scholar]
  13. Kogure K., Simidu U., Taga N. A tentative direct microscopic method for counting living marine bacteria. Can J Microbiol. 1979 Mar;25(3):415–420. doi: 10.1139/m79-063. [DOI] [PubMed] [Google Scholar]
  14. Meyer-Reil L. A. Autoradiography and epifluorescence microscopy combined for the determination of number and spectrum of actively metabolizing bacteria in natural water. Appl Environ Microbiol. 1978 Sep;36(3):506–512. doi: 10.1128/aem.36.3.506-512.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SILVERMAN M. P., LUNDGREN D. G. Studies on the chemoautotrophic iron bacterium Ferrobacillus ferrooxidans. I. An improved medium and a harvesting procedure for securing high cell yields. J Bacteriol. 1959 May;77(5):642–647. doi: 10.1128/jb.77.5.642-647.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schmidt E. L., Bakole R. O., Bohlool B. B. Fluorescent-antibody approach to study of rhizobia in soil. J Bacteriol. 1968 Jun;95(6):1987–1992. doi: 10.1128/jb.95.6.1987-1992.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Strayer R. F., Tiedje J. M. Application of the fluorescent-antibody technique to the study of a methanogenic bacterium in lake sediments. Appl Environ Microbiol. 1978 Jan;35(1):192–198. doi: 10.1128/aem.35.1.192-198.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ward T. E., Frea J. I. Sediment distribution of methanogenic bacteria in lake erie and cleveland harbor. Appl Environ Microbiol. 1980 Mar;39(3):597–603. doi: 10.1128/aem.39.3.597-603.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zimmermann R., Iturriaga R., Becker-Birck J. Simultaneous determination of the total number of aquatic bacteria and the number thereof involved in respiration. Appl Environ Microbiol. 1978 Dec;36(6):926–935. doi: 10.1128/aem.36.6.926-935.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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