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Identification and quantification of Thiothrix eikelboomii using qPCR for early detection of bulking incidents in a full-scale water reclamation plant

  • Environmental biotechnology
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Abstract

This study found that the ratio of Thiothrix eikelboomii to total bacterial concentrations (TH/TB) (%) was a better indicator of bulking incidents affecting effluent quality compared to absolute T. eikelboomii abundance alone. This was determined using a genus-specific Thiothrix quantitative PCR primer and probe set, which was developed in this study to monitor specific Thiothrix populations over a 1-year period. T. eikelboomii was identified as the source of bulking incidents based on sequencing of the 16S rRNA gene at a nitrifying–denitrifying wastewater treatment plant. Peak T. eikelboomii concentrations observed in March, April, and July 2009 were 2.32 × 1010, 2.64 × 1010, and 1.84 × 1010 cells/l, respectively. The highest fraction of T. eikelboomii to total bacterial population was measured at 0.24% in March, and a ratio >0.19% caused increases of suspended solids and biochemical oxygen demand in the secondary effluent. Additionally, food/mass ratios, dissolved oxygen concentrations in the anoxic selector, and ammonium ion concentrations in the primary effluent were three parameters displaying statistically significant correlations (r = 0.40, r = 0.50, and r = 0.32, respectively) to Thiothrix spp. abundance in an aeration tank. No bulking events caused by T. eikelboomii occurred when the dissolved oxygen concentrations in the anoxic selector was maintained at lower than 0.12 mg/l and the TH/TB ratios were <0.10%.

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Acknowledgments

We are thankful to Irvine Ranch Water District for laboratory personnel and operators at Michelson Water Reclamation Plant for their assistance with data collection, sample processing, and chemical analysis. This research was supported by the National Water Research Institute (Award number no. 08-TM.004), CA and US National Science Foundation (Fellowship Award 1015730). We acknowledge Dr. Linda Tseng for her assistance with MANOVA analysis and Dr. Tom Wang for cloning T. eikelboomii PCR amplicons. We would like to also acknowledge Fabricio Berger de Vargas and Andre Borin Venturini for performing the melt-curve analysis for all environmental samples.

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Authors and Affiliations

  1. Department of Civil Engineering and Construction Engineering Management, California State University, Long Beach, Long Beach, CA, 90840, USA

    Pitiporn Asvapathanagul

  2. Department of Civil and Environmental Engineering, University of California, Irvine, Irvine, CA, 92697, USA

    Betty H Olson

  3. Department of Civil and Environmental Engineering, University of California, Los Angeles, Los Angeles, CA, 90095, USA

    Phillip B Gedalanga

  4. Civil and Environmental Engineering, University of California, Los Angeles, USA

    Ardalan Hashemi

  5. Department of Environment and Engineering, Nan**g University of Science and Technology, Nan**g, 210094, China

    Zhonghua Huang

  6. Irvine Ranch Water District, Irvine, CA, 92612, USA

    Jason La

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  1. Pitiporn Asvapathanagul
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Correspondence to Betty H Olson.

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Asvapathanagul, P., Olson, B.H., Gedalanga, P.B. et al. Identification and quantification of Thiothrix eikelboomii using qPCR for early detection of bulking incidents in a full-scale water reclamation plant. Appl Microbiol Biotechnol 99, 4045–4057 (2015). https://doi.org/10.1007/s00253-014-6230-5

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