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Using qPCR to diagnose the problem after losing nitrification

11/19/2024

 
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Nitrification is one of the most complex and crucial processes within the wastewater microbial community. This biochemical process involves the conversion of ammonia (NH₃) into nitrate (NO₃⁻) and is carried out by a specialized group of microorganisms known as nitrifiers. These nitrifiers can be broadly categorized into two main groups: Ammonia Oxidizing Bacteria (AOB) and Nitrite Oxidizing Bacteria (NOB). Both are slow-growing organisms and require specific environmental conditions to thrive, including relatively narrow ranges of pH, alkalinity, and dissolved oxygen levels.

The Process of Nitrification
Nitrification occurs in two main stages:
  • Ammonia Oxidation: In the first stage, ammonia is oxidized to nitrite (NO₂⁻) by AOB. This step is represented by the chemical equation (note the following equations not balanced):
  • NH₃ + O₂ → NO₂⁻ + H₂O + H⁺
  • This reaction is primarily carried out by bacteria from genera such as Nitrosomonas, Nitrosospira, and Nitrosococcus.
  • Nitrite Oxidation: In the second stage, nitrite is further oxidized to nitrate by NOB. The chemical equation for this reaction is:
  • NO₂⁻ + O₂ → NO₃⁻
  • Common genera involved in this process include Nitrobacter, Nitrospira, and Nitrococcus.

Environmental Conditions for Optimal Nitrification
The efficiency of nitrification is highly dependent on several environmental factors:
  • pH
    ​
    Nitrifiers operate best within a narrow pH range of 6.5 to 8.5. Deviations outside this range can inhibit their activity and slow down the nitrification process. Acidic or highly alkaline conditions can adversely affect enzyme activity and cellular processes in these bacteria.
  • Alkalinity
    Alkalinity acts as a buffer to maintain a stable pH in the system. During nitrification, hydrogen ions (H⁺) are produced, which can lower the pH. Adequate alkalinity is necessary to neutralize these acids and maintain a conducive environment for nitrifiers.
  • Dissolved Oxygen
    Both AOB and NOB require dissolved oxygen (DO) for their metabolic processes. Optimal DO levels for nitrification typically range from 2 to 4 mg/L. Insufficient oxygen can lead to incomplete nitrification and the accumulation of intermediate compounds such as nitrite.

Challenges in Nitrification 
Inhibition and Activity Reduction
Facilities often report instances where they believe their nitrifiers have been "killed." However, in many cases, it is more likely that the nitrifiers are experiencing inhibition or a decrease in activity rather than a complete die-off. Factors such as the presence of toxic compounds, sudden changes in environmental conditions, or competition with other microorganisms can inhibit nitrification.

Monitoring and Diagnosis
Before implementing widespread changes, importing sludge from another plant, or adding concentrated nitrifier products, it is crucial to accurately diagnose the state of the nitrifier population. Aster Bio’s qPCR technology offers a powerful tool for this purpose. By quantifying the genetic material of nitrifiers, qPCR provides results within hours rather than days, enabling informed decision-making and reducing the need for costly and time-consuming interventions.

Benefits of qPCR Technology
  • Accurate & Rapid Results
    Traditional methods for assessing nitrifier populations, such as culture-based techniques, can take several days to yield results. qPCR technology, on the other hand, delivers rapid and accurate data, allowing for timely adjustments to the treatment process.
  • Cost and Time Efficiency
    By providing quick insights into the health and abundance of nitrifiers, qPCR helps facilities avoid unnecessary expenses associated with overhauling treatment processes or importing sludge. This technology enables operators to make targeted interventions based on real-time data, saving both time and money.

We Can Help
Are you struggling with nitrification in your wastewater treatment plant? Don't let uncertainty slow you down at a cost of $225 per sample, qPCR maintains permit and saves money. Contact Aster Bio’s experts today to learn how our qPCR technology can provide you with the rapid and accurate insights you need to optimize your nitrifier populations and enhance the efficiency of your treatment process. Let us help you save time, reduce costs, and achieve better outcomes for your facility. Contact us for more details at [email protected].

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    Author

    Erik Rumbaugh has been involved in biological waste treatment for over 20 years. He has worked with industrial and municipal wastewater  facilities to ensure optimal performance of their treatment systems. He is a founder of Aster Bio (www.asterbio.com) specializing in biological waste treatment.

    View my profile on LinkedIn

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