Journal of Oceanography

, Volume 62, Issue 6, pp 793–799 | Cite as

Comparison of bromodeoxyuridine immunoassay with tritiated thymidine radioassay for measuring bacterial productivity in oceanic waters

  • Koji Hamasaki


Recently, bromodeoxyuridine (BrdU) has been successfully applied to the measurement of bacterial productivity as an alternative to tritiated thymidine (3H-TdR), which is widely used but often restricted by regulations, particularly in field settings. Here, I report improvements to existing BrdU methods to simplify procedures and increase sensitivity. The feasibility of the method was tested measuring bacterial production in low-productive waters. The method provided radioisotope-free measurements of bacterial production rates at shorter (∼1 h) on-board processing time of samples than previously reported procedures. It was applicable to the detection of rates ranging from 0.021 to 2.7 pmol BrdU l−1h−1. BrdU incorporation rates measured by immunoassay showed a statistically significant correlation with 3H-TdR incorporation rates measured by radioassay (r = 0.74, n = 24, p < 0.001). The linear regression obtained (BrdU = 0.80[3H-TdR] − 0.016) showed a similar relationship to previously reported regressions (BrdU = 0.65[3H-TdR] + 0.12, [3H-BrdU] = 0.69[3H-TdR] − 0.81). There were no statistically significant differences among these regression lines. These results suggest that the method described here provides a non-radioisotopic productivity measurement of bacteria in oceanic epipelagic waters, while retaining continuity of the data with other existing 3H-TdR and BrdU methods.


Bacterial production thymidine bromodeoxyuridine 


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  1. Artursson, V. and J. K. Jansson (2003): Use of bromodeoxyuridine immunocapture to identify active bacteria associated with arbuscular mycorrhizal hyphae. Appl. Environ. Microb., 69, 6208–6215.CrossRefGoogle Scholar
  2. Azam, F. (1998): Microbial control of oceanic carbon flux: The plot thickens. Science, 280, 694–696.CrossRefGoogle Scholar
  3. Borneman, J. (1999): Culture-independent identification o microorganisms that respond to specific stimuli. Appl. Environ. Microbiol., 65, 3398–3400.Google Scholar
  4. Ducklow, H. (2000): Bacterial production and biomass in the oceans. p. 85–120. In Microbial Ecology of the Oceans, ed. by D. Kirchman and P. J. B. Williams, Wiley-Liss, New York.Google Scholar
  5. Ducklow, H. W. and C. A. Carlson (1992): Oceanic bacterial production. p. 113–181. In Advances in Microbial Ecology, ed. by K. C. Marshall, Plenum Press, New York.CrossRefGoogle Scholar
  6. Fuhrman, J. A. and F. Azam (1980): Bacterioplankton secondary production estimates for coastal waters of British Columbia, Antarctica and California. Appl. Environ. Microbiol., 39, 1085–1095.Google Scholar
  7. Fuhrman, J. A. and F. Azam (1982): Thymidine incorporation as a measure of heterotrohic bacterioplankton production in marine surface waters: evaluation and field results. Mar. Biol., 66, 109–120.CrossRefGoogle Scholar
  8. Giovannoni, S. J. and U. Sting (2005): Molecular diversity and ecology of microbial plankton. Nature, 437, 343–348.CrossRefGoogle Scholar
  9. Hamasaki, K., R. A. Long and F. Azam (2004): Individual cell growth rates of marine bacteria, measured by bromodeoxyuridine incorporation. Aquat. Microb. Ecol., 35, 217–227.CrossRefGoogle Scholar
  10. Hollibaugh, J. T. (1988): Limitation of the [3H]thymidine method for estimating bacterial productivity due to thymidine metabolism. Mar. Ecol. Prog. Ser., 43, 19–30.CrossRefGoogle Scholar
  11. Karner, M. B., E. F. DeLong and D. M. Karl (2001): Archeal dominance in the mesopelagic zone of the Pacific Ocean. Nature, 409, 507–510.CrossRefGoogle Scholar
  12. Kirchman, D. L., E. K’nees and R. Hodson (1985): Leucine incorporation and its potential as a measure of protein synthesis by bacteria in natural waters. Appl. Environ. Microbiol., 49, 599–607.Google Scholar
  13. Moriarty, D. J. W. (1986): Measurement of bacterial growth rates in aquatic systems from rates of nucleic acid synthesis. p. 245–292. In Advances in Microbial Ecology, ed. by K. C. Marshall, Plenum Press, New York.CrossRefGoogle Scholar
  14. Nelson, C. E. and C. A. Carlson (2005): A nonradioactive assay of bacterial productivity optimized for oligotrophic pelagic environments. Limnol. Oceanogr. Methods, 3, 211–220.CrossRefGoogle Scholar
  15. Pernthaler, A. and J. Pernthaler (2005): Diurnal variation of cell proliferation in three bacterial taxa from coastal North Sea waters. Appl. Environ. Microbiol., 71, 4638–4644.CrossRefGoogle Scholar
  16. Pernthaler, A., J. Pernthaler, M. Schattenhofer and R. Amann (2002): Identification of DNA-synthesizing bacterial cells in coastal North Sea plankton. Appl. Environ. Microbiol., 68, 5728–5736.CrossRefGoogle Scholar
  17. Robarts, R. D. and T. Zohary (1993): Fact or fiction-bacterial growth rates and production as determined by [methyl-3H]-thymidine? p. 371–425. In Advances in Microbial Ecology, ed. by J. G. Jones, Plenum Press, New York.CrossRefGoogle Scholar
  18. Sokal, R. R. and F. J. Rohlf (1995): Biometry. W.H. Freeman and Company, New York, 887 pp.Google Scholar
  19. Steward, G. and F. Azam (1999): Bromodeoxyuridine as an alternative to 3H-thymidine for measuring bacterial productivity in aquatic samples. Aquat. Microb. Ecol., 19, 57–66.CrossRefGoogle Scholar
  20. Suzuki, R. and T. Ishimaru (1990): An improved method for the determination of phytoplankton chlorophyll using N,N-dimethylformamide. J. Oceanogr., 46, 190–194.Google Scholar
  21. Urbach, E., K. L. Vergin and S. J. Giovanoni (1999): Immunochemical detection and isolation of DNA from metabolically active bacteria. Appl. Environ. Microbiol., 65, 1207–1213.Google Scholar
  22. Warnecke, F., R. Sommaruga, R. Sekar, J. S. Hofer and J. Pernthaler (2005): Abundance, identity, and growth state of Actinobacteria in mountain lakes of different UV transparency. Appl. Environ. Microbiol., 71, 5551–5559.CrossRefGoogle Scholar
  23. Yin, B., D. Crowley, G. Sparovek, W. J. De Melo and J. Borneman (2000): Bacterial functional redundancy along a soil reclamation gradient. Appl. Environ. Microbiol., 66, 4361–4365.CrossRefGoogle Scholar

Copyright information

© The Oceanographic Society of Japan/TERRAPUB/Springer 2006

Authors and Affiliations

  • Koji Hamasaki
    • 1
  1. 1.Graduate School of Biosphere ScienceHiroshima UniversityKagamiyama, Higashi-Hiroshima, HiroshimaJapan

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