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Related Experiment Videos

Solutions to problems in enumerating sediment bacteria by direct counts.

M Schallenberg1, J Kalff, J B Rasmussen

  • 1Department of Biology, McGill University, 1205 Ave. Dr. Penfield, Montreal, Quebec H3A 1B1, Canada.

Applied and Environmental Microbiology
|May 1, 1989
PubMed
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Accurate bacterial counts in sediments require optimal fluorochrome concentrations and corrections for particle masking. This study reveals a method to standardize DAPI staining for reliable sediment bacteria quantification.

Area of Science:

  • Environmental Microbiology
  • Microbial Ecology
  • Analytical Chemistry

Background:

  • Accurate quantification of bacteria in environmental samples like sediments is crucial for understanding microbial ecology.
  • The fluorochrome DAPI (4'-6-diamidino-2-phenylindole dihydrochloride) is widely used for bacterial enumeration.
  • Sediment properties can interfere with DAPI staining efficiency and introduce bias in bacterial counts.

Purpose of the Study:

  • To investigate the impact of sediment type on DAPI staining effectiveness and bacterial abundance.
  • To determine optimal DAPI concentrations for accurate bacterial enumeration in diverse sediments.
  • To develop a method to correct for sediment particle masking of DAPI-stained bacteria.

Main Methods:

  • Examined DAPI staining effectiveness across various concentrations and sediment types.

Related Experiment Videos

  • Assessed sediment particle masking effects on DAPI-stained bacteria.
  • Utilized turbidity (A(750)) and quartz-corrected water content to quantify masking and initial dilution.
  • Developed a correction procedure based on integrated dilution.

    • Main Results:

    • Sediment type significantly influences DAPI staining efficiency; many studies likely underestimate bacterial numbers.
    • A DAPI concentration of 5 µg/mL is recommended for effective staining in most gravity-core sediments.
    • A linear relationship between masking and integrated sediment dilution was established.
    • The developed correction method enables accurate and unbiased bacterial counts, revealing a strong correlation between bacteria and sediment organic matter.

    Conclusions:

    • The proposed method of DAPI staining with a correction for sediment masking allows for standardized and reliable quantification of sediment bacteria.
    • Accurate bacterial counts are essential for understanding the relationship between microbial communities and environmental factors like organic matter.
    • This approach provides a basis for consistent interpretation of bacterial abundance data across different sediment studies.