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. 1980 Apr;39(4):913–918. doi: 10.1128/aem.39.4.913-918.1980

Immunofluorescent Assay for the Marine Ammonium-Oxidizing Bacterium Nitrosococcus oceanus†

B B Ward 1, M J Perry 1
PMCID: PMC291443  PMID: 16345557

Abstract

Nitrification is one of the important microbiological transformations of nitrogen in the ocean. Traditional enrichment-culture methods for enumerating the autotrophic bacteria which oxidize ammonium to nitrite are very time consuming (months) and are believed to seriously underestimate natural abundances. A fluorescent-antibody assay for a marine ammonium-oxidizing bacterium was developed to provide a rapid and direct means of identifying these microorganisms. Antibodies to Nitrosococcus oceanus were prepared and tested against pure cultures of marine, freshwater, and soil ammonium oxidizers and against bacteria from natural seawater samples. Cell counts of culture samples determined by the fluorescent-antibody assay agreed with hemacytometer and acridine orange counts. Our results demonstrated that the immunofluorescent assay is a powerful tool for the detection of Nitrosococcus in the marine environment.

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Selected References

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  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. Belser L. W., Schmidt E. L. Serological diversity within a terrestrial ammonia-oxidizing population. Appl Environ Microbiol. 1978 Oct;36(4):589–593. doi: 10.1128/aem.36.4.589-593.1978. [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. CARLUCCI A. F., PRAMER D. Factors influencing the plate method for determining abundance of bacteria in sea water. Proc Soc Exp Biol Med. 1957 Nov;96(2):392–394. doi: 10.3181/00379727-96-23487. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Haaijman J. J., Bloemmen F. J., Ham C. M. Microfluorometric immunoassays with antigens bound to sepharose beads. Ann N Y Acad Sci. 1975 Jun 30;254:137–150. doi: 10.1111/j.1749-6632.1975.tb29163.x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Kenny G. E. Immunogenicity of Mycoplasma pneumoniae. Infect Immun. 1971 Apr;3(4):510–515. doi: 10.1128/iai.3.4.510-515.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Martinsen C. S., Zachariah P. K. Growth of various bacteria on polycyclic aromatic hydrocarbons and N-2-fluorenylacetamide. J Appl Bacteriol. 1978 Jun;44(3):365–371. doi: 10.1111/j.1365-2672.1978.tb00810.x. [DOI] [PubMed] [Google Scholar]
  11. Reed W. M., Dugan P. R. Distribution of Methylomonas methanica and Methylosinus trichosporium in Cleveland Harbor as Determined by an Indirect Fluorescent Antibody-Membrane Filter Technique. Appl Environ Microbiol. 1978 Feb;35(2):422–430. doi: 10.1128/aem.35.2.422-430.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Soriano S., Walker N. Isolation of ammonia-oxidizing autotrophic bacteria. J Appl Bacteriol. 1968 Dec;31(4):493–497. doi: 10.1111/j.1365-2672.1968.tb00397.x. [DOI] [PubMed] [Google Scholar]
  14. VISHNIAC W., SANTER M. The thiobacilli. Bacteriol Rev. 1957 Sep;21(3):195–213. doi: 10.1128/br.21.3.195-213.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]

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