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Applied and Environmental Microbiology, September 2009, p. 5555-5562, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.00407-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Monochloramine Disinfection Kinetics of Nitrosomonas europaea by Propidium Monoazide Quantitative PCR and Live/Dead BacLight Methods{triangledown}

David G. Wahman, Karen A. Wulfeck-Kleier, and Jonathan G. Pressman*

United States Environmental Protection Agency, Office of Research and Development, Cincinnati, Ohio

Received 18 February 2009/ Accepted 21 June 2009

Monochloramine disinfection kinetics were determined for the pure-culture ammonia-oxidizing bacterium Nitrosomonas europaea (ATCC 19718) by two culture-independent methods, namely, Live/Dead BacLight (LD) and propidium monoazide quantitative PCR (PMA-qPCR). Both methods were first verified with mixtures of heat-killed (nonviable) and non-heat-killed (viable) cells before a series of batch disinfection experiments with stationary-phase cultures (batch grown for 7 days) at pH 8.0, 25°C, and 5, 10, and 20 mg Cl2/liter monochloramine. Two data sets were generated based on the viability method used, either (i) LD or (ii) PMA-qPCR. These two data sets were used to estimate kinetic parameters for the delayed Chick-Watson disinfection model through a Bayesian analysis implemented in WinBUGS. This analysis provided parameter estimates of 490 mg Cl2-min/liter for the lag coefficient (b) and 1.6 x 10–3 to 4.0 x 10–3 liter/mg Cl2-min for the Chick-Watson disinfection rate constant (k). While estimates of b were similar for both data sets, the LD data set resulted in a greater k estimate than that obtained with the PMA-qPCR data set, implying that the PMA-qPCR viability measure was more conservative than LD. For N. europaea, the lag phase was not previously reported for culture-independent methods and may have implications for nitrification in drinking water distribution systems. This is the first published application of a PMA-qPCR method for disinfection kinetic model parameter estimation as well as its application to N. europaea or monochloramine. Ultimately, this PMA-qPCR method will allow evaluation of monochloramine disinfection kinetics for mixed-culture bacteria in drinking water distribution systems.

* Corresponding author. Mailing address: United States Environmental Protection Agency, 26 W. Martin Luther King Dr., Cincinnati, OH 45268. Phone: (513) 569-7625. Fax: (513) 487-2543. E-mail: pressman.jonathan{at}epa.gov

{triangledown} Published ahead of print on 26 June 2009.

Applied and Environmental Microbiology, September 2009, p. 5555-5562, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.00407-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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