Bacterial Growth and Productivity

  • Robert G. Wetzel
  • Gene E. Likens


Earlier discussions emphasized that dead organic matter, called detritus, exists as a spectrum from dissolved organic compounds, organic colloids, and larger particles of organic matter. Dissolved organic matter is in much greater abundance, by about five to ten times, than is particulate organic matter. All microflora must degrade particulate organic matter enzymati-cally to the dissolved form prior to assimilation for further metabolic breakdown and eventual mineralization to inorganic solutes and gases.

The rates of growth by bacteria and their productivity are known poorly from aquatic ecosystems. Much of this lack of progress resulted from the slow application of laboratory methodology to heterogeneous natural communities of bacteria. Often methods were developed for isolated strains at high laboratory concentrations. A number of methods have been developed recently that permit examination of growth and productivity of in situ communities of bacteria. These methods are evolving as more is learned about metabolic constraints and variations.

Natural variance is high in microbial communities, and many organisms are metabolically inactive or dormant. The methods used to examine bacteria are labor intensive. We describe methods here in considerable detail because they require rigorous application in order to obtain meaningful results. Conversion of rates of biomass change or genetic replication to actual rates of carbon flux in bacterial secondary productivity is difficult and is undergoing intensive study by many researchers. A spectrum of conversion factors is given for different environmental conditions in order to indicate the range of probable in situ values.

We stress the importance of these evaluations of in situ metabolism and growth rates of bacteria. Much of the understanding about biogeo-chemical cycling in aquatic ecosystem has been limited severely by poor or inadequate techniques for evaluation of bacterial metabolism and its limitations by environmental parameters. The methods presented here are not perfect, and they are laborious. Certainly they will improve with time. However, these methods offer means to examine in situ rates of bacterial growth and productivity and, importantly, means to evaluate in situ effects of experimental manipulations of environmental conditions.


Acridine Orange Thymidine Incorporation Tritiated Thymidine Lewis Publisher Thymidine Uptake 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Austin, B. (ed.). 1988. Methods in Aquatic Bacteriology. Wiley, Chichester, 425 pp.Google Scholar
  2. Autio, R.M. 1990. Bacterioplankton in filtered brackish water cultures: Some physical and chemical parameters affecting community properties. Arch. Hydrobiol. 117:437–451.Google Scholar
  3. Bell, R.T., G.M. Ahlgren, and I. Ahlgren. 1983. Estimating bacterioplankton production by the [3H]thymidine incorporation in a eutrophic Swedish lake. Appl. Environ. Microbiol. 45:1709–1721.PubMedGoogle Scholar
  4. Bowden, W.B. 1977. Comparison of two direct-count techniques for enumerating aquatic bacteria. Appl. Environ. Microbiol. 33:1229–1232.PubMedGoogle Scholar
  5. Bratbak, G. 1985. Bacterial biovolume and biomass estimations. Appl. Environ. Microbiol. 49:1488–1493.PubMedGoogle Scholar
  6. Bratbak, G. 1993. Microscope methods for measuring bacterial biovolume: Epifluorescence microscopy, scanning electron microscopy, and transmission electron microscopy, pp. 309–317. In: P.F. Kemp, B.F. Sherr, E.B. Sherr, and J.J. Cole, Editors. Handbook of Methods in Aquatic Microbial Ecology. Lewis Publishers, Boca Raton.Google Scholar
  7. Bratbak, G. and I. Dundas. 1984. Bacterial dry matter content and biomass estimations. Appl. Environ. Microbiol. 48:155–151.Google Scholar
  8. Bührer, H. 1977. Verbesserte Acridinorangemethode zur Direktzählung von Bakterien aus Seesediment. Schweiz. Z. Hydrol. 39:99–103.Google Scholar
  9. Button, D.K. and B.R. Robertson. 1993. Use of high-resolution flow cytometry to determine the activity and distribution of aquatic bacteria, pp. 163–173. In: P.F. Kemp, B.F. Sherr, E. B. Sherr, and J. J. Cole, Editors. Handbook of Methods in Aquatic Microbial Ecology. Lewis Publishers, Boca Raton.Google Scholar
  10. Caldwell, D.E. 1977. The planktonic microflora of lakes. CRC Critical Rev. Microbiol. 5:305–370.Google Scholar
  11. Coveney, M.F and R.G. Wetzel. 1988. Experimental evaluation of conversion factors for the (3H)thymidine incorporation assay of bacterial secondary productivity. Appl. Environ. Microbiol. 54:2018–2026.PubMedGoogle Scholar
  12. Coveney, M.F. and R.G. Wetzel. 1989. Bacterial metabolism of algal extracellular carbon. Hydrobiologia 173:141–149.CrossRefGoogle Scholar
  13. Daley, R.J. 1979. Direct epifluorescence enumeration of native aquatic bacteria: Uses, limitations, and comparative accuracy, pp 29–45. In: J.W Costerton and R.R. Colwell, Editors. Native Aquatic Bacteria: Enumeration, Activity, and Ecology. ASTM STP695. American Society of Testing Materials, Washington, D.C.Google Scholar
  14. Ducklow, H.W. and C.A. Carlson. 1992. Oceanic bacterial production. Adv. Microbial Ecol. 12:113–181.CrossRefGoogle Scholar
  15. Ducklow, H.W. and S.M. Hill. 1985. Tritiated thymidine incorporation and the growth of heterotrophic bacteria in warm core rings. Limnol. Oceanogr. 30:260–272.CrossRefGoogle Scholar
  16. van Es, F.B. and L.-A. Meyer-Reil. 1983. Biomass and metabolic activity of heterotrophic marine bacteria. Adv. Microbial Ecol. 6:111–170.Google Scholar
  17. Francisco, D.E., R.A. Mah, and A.C. Rabin. 1973. Acridine orange-epifluorescence technique for counting bacteria in natural waters. Trans. Amer. Microsc. Soc. 92:416–421.CrossRefGoogle Scholar
  18. Fuhrman, J.A. and F. Azam. 1982. Thymidine incorporation as a measure of heterotrophic bacterioplankton production in marine surface waters: Evaluation and field results. Mar. Biol. 66:109–120.CrossRefGoogle Scholar
  19. Hobbie, J.E., R.J. Daley, and S. Jasper. 1977. Use of Nuclepore filters for counting bacteria by fluorescence microscopy. Appl. Environ. Microbiol. 33:1225–1228.PubMedGoogle Scholar
  20. Kemp, P.F., B.F. Sherr, E.B. Sherr, and J.J. Cole (eds). 1993. Handbook of Methods in Aquatic Microbial Ecology. Lewis Publishers, Boca Raton. 777 pp.Google Scholar
  21. Kirchman, D.L. 1993. Leucine incorporation as a measure of biomass production by heterotrophic bacteria, pp. 509–512. In: P.F. Kemp, B.F. Sherr, E.B. Sherr, and J.J. Cole, Editors. Handbook of Methods in Aquatic Microbial Ecology. Lewis Publishers, Boca RatonGoogle Scholar
  22. Kirchman, D., H. Ducklow, and R. Mitchell. 1982. Estimates of bacterial growth from changes in uptake rates and biomass. Appl. Environ. Microbiol. 44:1296–1307.PubMedGoogle Scholar
  23. Kirchman, D.L., E. K’Nees, and R. Hudson. 1985. Leucine incorporation and its potential as a measure of protein synthesis by bacteria in natural aquatic systems. Appl. Environ. Microbiol. 49:599–601.PubMedGoogle Scholar
  24. Kirchman, D.L., S.Y. Newell, and R.E. Hodson. 1986. Incorporation versus biosynthesis of leucine: Implications for measuring rates of protein synthesis and biomass production by bacteria in marine systems. Mar. Ecol. Prog. Ser. 32:47–59.CrossRefGoogle Scholar
  25. Lovell, C.R. and A. Konopka. 1985. Seasonal bacterial production in a dimictic lake as measured by increases in cell numbers and thymidine incorporation. Appl. Environ. Microbiol. 49:492–500.PubMedGoogle Scholar
  26. Moriarty, D.J.W. 1986. Measurement of bacterial growth rates in aquatic systems using rates of nucleic acid synthesis. Adv. Aquatic Microbiol. 9:245–292.Google Scholar
  27. Moriarty, D.J.W. 1989. Accurate conversion factors for calculating bacterial growth rates from thymidine incorporation into DNA: Elusive or illusive? Ergebn. Limnol. Arch. Hydrobiol. (In press).Google Scholar
  28. Murray, R.E. and R.E. Hodson. 1985. Annual cycle of bacterial secondary production in five aquatic habitats of the Okefenokee Swamp ecosystem. Appl. Environ. Microbiol. 49:650–655.PubMedGoogle Scholar
  29. Norland, S. 1993. The relationship between biomass and volume of bacteria, pp. 303–307. In: P.F. Kemp, B.F. Sherr, E.B. Sherr, and J.J. Cole, Editors. Handbook of Methods in Aquatic Microbial Ecology. Lewis Publishers, Boca Raton.Google Scholar
  30. Pomeroy, L.R. 1974. The ocean’s food web: A changing paradigm. BioScience 9:499–504.CrossRefGoogle Scholar
  31. Porter, K.G. and Y.S. Feig. 1980. The use of DAPI for identifying and counting aquatic microflora. Limnol. Oceanogr. 25:943–948.CrossRefGoogle Scholar
  32. Psenner, R. 1990. From image analysis to chemical analysis of bacteria: A long-term study? Limnol. Oceanogr. 35:234–237.Google Scholar
  33. Riemann, B. and M. Sjøndergaard. 1985. Regulation of bacterial secondary production in two eutrophic lakes and in experimental enclosures. J. Plankton Res. 8:519–536.CrossRefGoogle Scholar
  34. Riemann, B., P.K. Bjørnsen, S. Newell, and R. Fallon. 1987. Calculation of cell production of coastal marine bacteria based on measured incorporation of [3H]thymidine. Limnol. Oceanogr. 32:471–476.CrossRefGoogle Scholar
  35. Robarts, R.D. and R.J. Wicks. 1989. [Methyl-3H]thymidine macromolecular incorporation and lipid labeling: Their significance to DNA labeling during measurements of aquatic bacterial growth rate. Limnol. Oceanogr. 34:213–222.CrossRefGoogle Scholar
  36. Robarts, R.D. and T. Zohary. 1993. Fact or fiction—Bacterial growth rates and production as determined by [methyl-3H]-thymidine? Adv. Microbial Ecol. 13:371–425.CrossRefGoogle Scholar
  37. Scavia, D. and G.A. Laird. 1987. Bacterioplankton in Lake Michigan: Dynamics, controls, and significance to carbon flux. Limnol. Oceanogr. 32:1017–1033.CrossRefGoogle Scholar
  38. Simon, M. and F. Azam. 1989. Protein content and protein synthesis rates of planktonic marine bacteria. Mar. Ecol. Prog. Ser. 51:201–213.CrossRefGoogle Scholar
  39. Sorokin, Y.I. and H. Kadota (eds). 1972. Techniques for the Assessment of Microbial Production and Decomposition in Fresh Waters. Blackwell, Oxford.Google Scholar

Copyright information

© Springer Science+Business Media New York 2000

Authors and Affiliations

  • Robert G. Wetzel
    • 1
  • Gene E. Likens
    • 2
  1. 1.Department of Biology, College of Arts and SciencesUniversity of AlabamaTuscaloosaUSA
  2. 2.Institute of Ecosystem Studies, Cary ArboretumThe New York Botanical GardenMillbrookUSA

Personalised recommendations