This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Karner, M.
Right arrow Articles by Fuhrman, J. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Karner, M.
Right arrow Articles by Fuhrman, J. A.
Agricola
Right arrow Articles by Karner, M.
Right arrow Articles by Fuhrman, J. A.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Apr 1997, 1208-1213, Vol 63, No. 4
Copyright © 1997, American Society for Microbiology

Determination of Active Marine Bacterioplankton: a Comparison of Universal 16S rRNA Probes, Autoradiography, and Nucleoid Staining

M Karner and JA Fuhrman
Department of Biological Sciences, Allan Hancock Foundation, University of Southern California, Los Angeles, California 90089-0371

We compared several currently discussed methods for the assessment of bacterial numbers and activity in marine waters, using samples from a variety of marine environments, from aged offshore seawater to rich harbor water. Samples were simultaneously tested for binding to a fluorescently labeled universal 16S rRNA probe; (sup3)H-labeled amino acid uptake via autoradiography; nucleoid-containing bacterial numbers by modified DAPI (4(prm1),6-diamidino-2-phenylindole) staining; staining with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), a compound supposed to indicate oxidative cell metabolism; and total bacterial counts (classical DAPI staining), taken as a reference. For the universal-probe counts, we used an image intensifying and processing system coupled to the epifluorescence microscope. All of the above-mentioned methods yielded lower cell counts than DAPI total counts. Universal-probe counts averaged about half of the corresponding DAPI count and were highly correlated to autoradiography counts (r(sup2) = 0.943; n = 7). Nucleoid-containing cell counts could be lower than DAPI counts by as much as 1 order of magnitude but sometimes matched autoradiography or probe counts. CTC counts were 2 orders of magnitude below DAPI counts. Universal 16S rRNA probe counts correlated well with autoradiography results, indicating a population with at least minimal metabolic activity. The greater variability of the nucleoid-containing cell counts calls for further investigation of the processes involved, and CTC counts were well below the range of the other methods tested.


This article has been cited by other articles:

  • Straza, T. R. A., Cottrell, M. T., Ducklow, H. W., Kirchman, D. L. (2009). Geographic and Phylogenetic Variation in Bacterial Biovolume as Revealed by Protein and Nucleic Acid Staining. Appl. Environ. Microbiol. 75: 4028-4034 [Abstract] [Full Text]  
  • Chavez de Paz, L. E., Hamilton, I. R., Svensater, G. (2008). Oral bacteria in biofilms exhibit slow reactivation from nutrient deprivation. Microbiology 154: 1927-1938 [Abstract] [Full Text]  
  • Falcioni, T., Papa, S., Gasol, J. M. (2008). Evaluating the Flow-Cytometric Nucleic Acid Double-Staining Protocol in Realistic Situations of Planktonic Bacterial Death. Appl. Environ. Microbiol. 74: 1767-1779 [Abstract] [Full Text]  
  • Eller, G., Tobe, K., Medlin, L. K. (2007). Hierarchical probes at various taxonomic levels in the Haptophyta and a new division level probe for the Heterokonta. J PLANKTON RES 29: 629-640 [Abstract] [Full Text]  
  • Lau, W. W. Y., Keil, R. G., Armbrust, E. V. (2007). Succession and Diel Transcriptional Response of the Glycolate-Utilizing Component of the Bacterial Community during a Spring Phytoplankton Bloom. Appl. Environ. Microbiol. 73: 2440-2450 [Abstract] [Full Text]  
  • Caro, A., Gros, O., Got, P., De Wit, R., Troussellier, M. (2007). Characterization of the Population of the Sulfur-Oxidizing Symbiont of Codakia orbicularis (Bivalvia, Lucinidae) by Single-Cell Analyses. Appl. Environ. Microbiol. 73: 2101-2109 [Abstract] [Full Text]  
  • Lin, X., Wakeham, S. G., Putnam, I. F., Astor, Y. M., Scranton, M. I., Chistoserdov, A. Y., Taylor, G. T. (2006). Comparison of Vertical Distributions of Prokaryotic Assemblages in the Anoxic Cariaco Basin and Black Sea by Use of Fluorescence In Situ Hybridization. Appl. Environ. Microbiol. 72: 2679-2690 [Abstract] [Full Text]  
  • Paoli, A., Karuza, A., de Vittor, C., del Negro, P., Umani, S. F. (2006). Daily variations of highly active bacteria in the Northern Adriatic Sea. J PLANKTON RES 28: 325-335 [Abstract] [Full Text]  
  • Elifantz, H., Malmstrom, R. R., Cottrell, M. T., Kirchman, D. L. (2005). Assimilation of Polysaccharides and Glucose by Major Bacterial Groups in the Delaware Estuary. Appl. Environ. Microbiol. 71: 7799-7805 [Abstract] [Full Text]  
  • Yokokawa, T., Nagata, T. (2005). Growth and Grazing Mortality Rates of Phylogenetic Groups of Bacterioplankton in Coastal Marine Environments. Appl. Environ. Microbiol. 71: 6799-6807 [Abstract] [Full Text]  
  • Pernthaler, J., Amann, R. (2005). Fate of Heterotrophic Microbes in Pelagic Habitats: Focus on Populations. Microbiol. Mol. Biol. Rev. 69: 440-461 [Abstract] [Full Text]  
  • Simu, K., Holmfeldt, K., Zweifel, U. L., Hagstrom, A. (2005). Culturability and Coexistence of Colony-Forming and Single-Cell Marine Bacterioplankton. Appl. Environ. Microbiol. 71: 4793-4800 [Abstract] [Full Text]  
  • Malmstrom, R. R., Cottrell, M. T., Elifantz, H., Kirchman, D. L. (2005). Biomass Production and Assimilation of Dissolved Organic Matter by SAR11 Bacteria in the Northwest Atlantic Ocean. Appl. Environ. Microbiol. 71: 2979-2986 [Abstract] [Full Text]  
  • Herndl, G. J., Reinthaler, T., Teira, E., van Aken, H., Veth, C., Pernthaler, A., Pernthaler, J. (2005). Contribution of Archaea to Total Prokaryotic Production in the Deep Atlantic Ocean. Appl. Environ. Microbiol. 71: 2303-2309 [Abstract] [Full Text]  
  • Malmstrom, R. R., Kiene, R. P., Cottrell, M. T., Kirchman, D. L. (2004). Contribution of SAR11 Bacteria to Dissolved Dimethylsulfoniopropionate and Amino Acid Uptake in the North Atlantic Ocean. Appl. Environ. Microbiol. 70: 4129-4135 [Abstract] [Full Text]  
  • Takai, K., Oida, H., Suzuki, Y., Hirayama, H., Nakagawa, S., Nunoura, T., Inagaki, F., Nealson, K. H., Horikoshi, K. (2004). Spatial Distribution of Marine Crenarchaeota Group I in the Vicinity of Deep-Sea Hydrothermal Systems. Appl. Environ. Microbiol. 70: 2404-2413 [Abstract] [Full Text]  
  • Lam, P., Cowen, J. P. (2004). Processing Deep-Sea Particle-Rich Water Samples for Fluorescence In Situ Hybridization: Consideration of Storage Effects, Preservation, and Sonication. Appl. Environ. Microbiol. 70: 25-33 [Abstract] [Full Text]  
  • Junge, K., Eicken, H., Deming, J. W. (2004). Bacterial Activity at -2 to -20{degrees}C in Arctic Wintertime Sea Ice. Appl. Environ. Microbiol. 70: 550-557 [Abstract] [Full Text]  
  • Burkert, U., Warnecke, F., Babenzien, D., Zwirnmann, E., Pernthaler, J. (2003). Members of a Readily Enriched {beta}-Proteobacterial Clade Are Common in Surface Waters of a Humic Lake. Appl. Environ. Microbiol. 69: 6550-6559 [Abstract] [Full Text]  
  • Nielsen, J. L., Aquino de Muro, M., Nielsen, P. H. (2003). Evaluation of the Redox Dye 5-Cyano-2,3-Tolyl-Tetrazolium Chloride for Activity Studies by Simultaneous Use of Microautoradiography and Fluorescence In Situ Hybridization. Appl. Environ. Microbiol. 69: 641-643 [Abstract] [Full Text]  
  • Bruns, A., Cypionka, H., Overmann, J. (2002). Cyclic AMP and Acyl Homoserine Lactones Increase the Cultivation Efficiency of Heterotrophic Bacteria from the Central Baltic Sea. Appl. Environ. Microbiol. 68: 3978-3987 [Abstract] [Full Text]  
  • Luna, G. M., Manini, E., Danovaro, R. (2002). Large Fraction of Dead and Inactive Bacteria in Coastal Marine Sediments: Comparison of Protocols for Determination and Ecological Significance. Appl. Environ. Microbiol. 68: 3509-3513 [Abstract] [Full Text]  
  • Pernthaler, A., Pernthaler, J., Amann, R. (2002). Fluorescence In Situ Hybridization and Catalyzed Reporter Deposition for the Identification of Marine Bacteria. Appl. Environ. Microbiol. 68: 3094-3101 [Abstract] [Full Text]  
  • Pernthaler, A., Preston, C. M., Pernthaler, J., DeLong, E. F., Amann, R. (2002). Comparison of Fluorescently Labeled Oligonucleotide and Polynucleotide Probes for the Detection of Pelagic Marine Bacteria and Archaea. Appl. Environ. Microbiol. 68: 661-667 [Abstract] [Full Text]  
  • Eilers, H., Pernthaler, J., Peplies, J., Glockner, F. O., Gerdts, G., Amann, R. (2001). Isolation of Novel Pelagic Bacteria from the German Bight and Their Seasonal Contributions to Surface Picoplankton. Appl. Environ. Microbiol. 67: 5134-5142 [Abstract] [Full Text]  
  • Sondergaard, M., Danielsen, M. (2001). Active bacteria (CTC+) in temperate lakes: temporal and cross-system variations. J PLANKTON RES 23: 1195-1206 [Abstract] [Full Text]  
  • Gregori, G., Citterio, S., Ghiani, A., Labra, M., Sgorbati, S., Brown, S., Denis, M. (2001). Resolution of Viable and Membrane-Compromised Bacteria in Freshwater and Marine Waters Based on Analytical Flow Cytometry and Nucleic Acid Double Staining. Appl. Environ. Microbiol. 67: 4662-4670 [Abstract] [Full Text]  
  • Pernthaler, A., Pernthaler, J., Eilers, H., Amann, R. (2001). Growth Patterns of Two Marine Isolates: Adaptations to Substrate Patchiness?. Appl. Environ. Microbiol. 67: 4077-4083 [Abstract] [Full Text]  
  • Lebaron, P., Servais, P., Agogué, H., Courties, C., Joux, F. (2001). Does the High Nucleic Acid Content of Individual Bacterial Cells Allow Us To Discriminate between Active Cells and Inactive Cells in Aquatic Systems?. Appl. Environ. Microbiol. 67: 1775-1782 [Abstract] [Full Text]  
  • Cottrell, M. T., Kirchman, D. L. (2000). Community Composition of Marine Bacterioplankton Determined by 16S rRNA Gene Clone Libraries and Fluorescence In Situ Hybridization. Appl. Environ. Microbiol. 66: 5116-5122 [Abstract] [Full Text]  
  • Ouverney, C. C., Fuhrman, J. A. (2000). Marine Planktonic Archaea Take Up Amino Acids. Appl. Environ. Microbiol. 66: 4829-4833 [Abstract] [Full Text]  
  • Gray, N. D., Howarth, R., Pickup, R. W., Jones, J. G., Head, I. M. (2000). Use of Combined Microautoradiography and Fluorescence In Situ Hybridization To Determine Carbon Metabolism in Mixed Natural Communities of Uncultured Bacteria from the Genus Achromatium. Appl. Environ. Microbiol. 66: 4518-4522 [Abstract] [Full Text]  
  • Cottrell, M. T., Kirchman, D. L. (2000). Natural Assemblages of Marine Proteobacteria and Members of the Cytophaga-Flavobacter Cluster Consuming Low- and High-Molecular-Weight Dissolved Organic Matter. Appl. Environ. Microbiol. 66: 1692-1697 [Abstract] [Full Text]  
  • DeLong, E. F., Taylor, L. T., Marsh, T. L., Preston, C. M. (1999). Visualization and Enumeration of Marine Planktonic Archaea and Bacteria by Using Polyribonucleotide Probes and Fluorescent In Situ Hybridization. Appl. Environ. Microbiol. 65: 5554-5563 [Abstract] [Full Text]  
  • Gray, N. D., Howarth, R., Pickup, R. W., Jones, J. G., Head, I. M. (1999). Substrate Uptake by Uncultured Bacteria from the Genus Achromatium Determined by Microautoradiography. Appl. Environ. Microbiol. 65: 5100-5106 [Abstract] [Full Text]  
  • Gasol, J. M., Zweifel, U. L., Peters, F., Fuhrman, J. A., Hagström, A. (1999). Significance of Size and Nucleic Acid Content Heterogeneity as Measured by Flow Cytometry in Natural Planktonic Bacteria. Appl. Environ. Microbiol. 65: 4475-4483 [Abstract] [Full Text]  
  • Sieracki, M. E., Cucci, T. L., Nicinski, J. (1999). Flow Cytometric Analysis of 5-Cyano-2,3-Ditolyl Tetrazolium Chloride Activity of Marine Bacterioplankton in Dilution Cultures. Appl. Environ. Microbiol. 65: 2409-2417 [Abstract] [Full Text]  
  • Ouverney, C. C., Fuhrman, J. A. (1999). Combined Microautoradiography-16S rRNA Probe Technique for Determination of Radioisotope Uptake by Specific Microbial Cell Types In Situ. Appl. Environ. Microbiol. 65: 1746-1752 [Abstract] [Full Text]  
  • Christensen, H., Hansen, M., Sørensen, J. (1999). Counting and Size Classification of Active Soil Bacteria by Fluorescence In Situ Hybridization with an rRNA Oligonucleotide Probe. Appl. Environ. Microbiol. 65: 1753-1761 [Abstract] [Full Text]  
  • Urbach, E., Vergin, K. L., Giovannoni, S. J. (1999). Immunochemical Detection and Isolation of DNA from Metabolically Active Bacteria. Appl. Environ. Microbiol. 65: 1207-1213 [Abstract] [Full Text]  
  • Jürgens, K., Pernthaler, J., Schalla, S., Amann, R. (1999). Morphological and Compositional Changes in a Planktonic Bacterial Community in Response to Enhanced Protozoan Grazing. Appl. Environ. Microbiol. 65: 1241-1250 [Abstract] [Full Text]  
  • Lisle, J. T., Broadaway, S. C., Prescott, A. M., Pyle, B. H., Fricker, C., McFeters, G. A. (1998). Effects of Starvation on Physiological Activity and Chlorine Disinfection Resistance in Escherichia coli O157:H7. Appl. Environ. Microbiol. 64: 4658-4662 [Abstract] [Full Text]  
  • Lebaron, P., Catala, P., Parthuisot, N. (1998). Effectiveness of SYTOX Green Stain for Bacterial Viability Assessment. Appl. Environ. Microbiol. 64: 2697-2700 [Abstract] [Full Text]  
  • Lebaron, P., Parthuisot, N., Catala, P. (1998). Comparison of Blue Nucleic Acid Dyes for Flow Cytometric Enumeration of Bacteria in Aquatic Systems. Appl. Environ. Microbiol. 64: 1725-1730 [Abstract] [Full Text]  
  • Weinbauer, M. G., Höfle, M. G. (1998). Significance of Viral Lysis and Flagellate Grazing as Factors Controlling Bacterioplankton Production in a Eutrophic Lake. Appl. Environ. Microbiol. 64: 431-438 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»