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Application of quantitative RT-PCR to determine the distribution of Microthrix parvicella in full-scale activated sludge treatment systems

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Abstract

Three wastewater treatment plants in South Africa were investigated to understand the phylogeny and distribution of Microthrix parvicella using real-time polymerase chain reaction (RT-PCR). The phylogenetic analysis of the 16S rRNA of M. parvicella revealed 98% to 100% homology of South African clones to M. parvicella reported in Genbank. The standard curves for RT-PCR showed R2 values greater than 0.99, accurate for quantification. The relative occurrence of M. parvicella 16S rRNA gene copies in the three wastewater treatment plants was in the range 0% to 3.97%. M. parvicella copies increased when the environmental temperature (≤20°C) and food/microorganism (F/M) ratio was low. The M. parvicella 16S rRNA copies could be positively correlated to the sludge volume index at low temperature. At higher temperature, there was a rapid reduction in M. parvicella population irrespective of other favorable factors, indicating the strong influence of temperature on filamentous proliferation. RT-PCR has potential applications in wastewater treatment plants to monitor sudden shift in the microbial population and assessing the plants efficacy.

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Acknowledgment

This research was financially supported by the National Research Foundation, South Africa and the Durban University of Technology.

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Correspondence to S. K. Sheena Kumari.

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Kumari, S.K.S., Marrengane, Z. & Bux, F. Application of quantitative RT-PCR to determine the distribution of Microthrix parvicella in full-scale activated sludge treatment systems. Appl Microbiol Biotechnol 83, 1135–1141 (2009). https://doi.org/10.1007/s00253-009-2013-9

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Keywords

  • Microthrix parvicella
  • 16S rRNA analysis
  • Real-time polymerase chain reaction
  • Food/microorganism ratio
  • Sludge volume index