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A comparative assessment of molecular biological and direct microscopic techniques for assessing aquatic systems

Environmental Monitoring and Assessment volume 145, pages 465–473 (2008)Cite this article

Abstract

Molecular techniques have clear value for community characterization; however, almost all previous datasets are based upon non-molecular measurements and it is hard to compare “old” data with “new” data because few correlations have been made. Therefore, the purpose of this evaluation was to simultaneously use molecular and non-molecular methods within the same sampling program to determine how data compare. Three methods were used for characterizing microbial populations in Lake 260 (L260) at Experimental Lakes Area (Ontario, Canada) during a whole-lake exposure study. Methods included whole-cell microscopic counts (for bacteria, cyanobacteria, algae and zooplankton), chlorophyll a, and small sub-unit (ssu)-rRNA hybridization using EUB-338, EUB-785, CYAN-785-a/b, EUCA-1379 and UNIV-1390 gene probes. Strong correlations were found between the EUB-785 probe signal and “bacteria minus cyanobacteria” direct counts, and the EUB-338 probe signal and “bacteria plus cyanobacteria” counts. Furthermore, the difference in probe signal between EUB-338 and EUB-785 (a presumptive signature for cyanobacteria and plastids) correlated with cyanobacterial direct counts and also with chlorophyll a. However, EUCA-1379 probe signal did not correlate with algae counts, and UNIV-1390 probe signal only correlated with total bacteria counts. Results suggest that, although ssu-rRNA methods are fast, reproducible, and specifically detect “viable” organisms, their use may be limited to non-eukaryotic populations unless new probes are developed that are more specific.

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Author notes

  1. K. A. Kidd

    Present address: Canadian Rivers Institute and Biology Department, University of New Brunswick, Saint John, New Brunswick, Canada

Authors and Affiliations

  1. Department of Civil, Environmental and Architectural Engineering, University of Kansas, Lawrence, Kansas, USA

    C. W. Knapp & D. W. Graham

  2. Fisheries and Oceans Canada, Freshwater Institute, Winnipeg, Manitoba, Canada

    D. L. Findlay & K. A. Kidd

  3. School of Civil Engineering and Geosciences, Cassie Building, Newcastle University, Newcastle Upon Tyne, NE1 7RU, UK

    C. W. Knapp

Authors
  1. C. W. Knapp
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  2. D. L. Findlay
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  3. K. A. Kidd
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  4. D. W. Graham
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Correspondence to C. W. Knapp.

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Knapp, C.W., Findlay, D.L., Kidd, K.A. et al. A comparative assessment of molecular biological and direct microscopic techniques for assessing aquatic systems. Environ Monit Assess 145, 465–473 (2008). https://doi.org/10.1007/s10661-007-0055-y

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  • DOI: https://doi.org/10.1007/s10661-007-0055-y

Keywords

  • 16S rRNA
  • Algae
  • Bacteria
  • Chlorophyll
  • Hybridization
  • Microscopy
  • Plastid
  • Whole-lake