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A rapid method to quantify nitrifiers in activated sludge.

Reto Manser1, Kathrin Muche, Willi Gujer

  • 1Swiss Federal Institute for Environmental Science and Technology (EAWAG), Zurich, Switzerland. reto.manser@eawag.ch

Water Research
|May 10, 2005
PubMed
Summary

A new, rapid method using Fluorescence In Situ Hybridization (FISH) and epifluorescence microscopy quickly quantifies nitrifying bacteria in activated sludge. This technique significantly reduces analysis time and effort compared to traditional methods.

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Area of Science:

  • Environmental microbiology
  • Wastewater treatment technologies

Background:

  • Accurate quantification of nitrifying bacteria is crucial for activated sludge process efficiency.
  • Traditional methods like FISH with confocal laser scanning microscopy (CLSM) are time-consuming and require expensive equipment.

Purpose of the Study:

  • To develop a rapid and cost-effective method for quantifying nitrifying bacteria in activated sludge.
  • To establish optimal parameters for manual counting using epifluorescence microscopy.

Main Methods:

  • Developed a method using Fluorescence In Situ Hybridization (FISH) and epifluorescence microscopy.
  • Quantified bacterial biovolume by manually counting aggregates and determining their size.
  • Evaluated method uncertainty based on the number of analyzed microscopic fields.

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Main Results:

  • Optimal field counts: 10-15 for ammonia-oxidizing bacteria, 6-8 for nitrite-oxidizing bacteria.
  • Reduced sample analysis time to 5-15 minutes from approximately 1 hour.
  • Demonstrated an explicit correlation between determined biovolume and measured bacterial activity.

Conclusions:

  • The developed FISH and epifluorescence microscopy method offers a rapid, accurate, and less resource-intensive alternative for nitrifying bacteria quantification.
  • This method enables efficient measurement of extended time series, aiding in process monitoring and optimization.
  • The strong correlation between biovolume and activity validates the method's utility in assessing microbial populations in activated sludge.