Development and application of real-time PCR for quantification of specific ammonia-oxidizing bacteria in activated sludge of sewage treatment systems

Applied Microbiology and Biotechnology
T LimpiyakornO Yagi

Abstract

In this study, four real-time polymerase chain reaction (PCR) primer sets were developed for the 16S rRNA genes of specific ammonia-oxidizing bacteria (AOB) found in activated sludge of sewage treatment systems. The primer sets target two of several sequence types of the Nitrosomonas oligotropha cluster, members within the Nitrosomonas communis cluster, and all members of the Nitrosomonas europaea-Nitrosococcus mobilis cluster. The detection limit of each primer set was in the range of 3x10(1)-6x10(2) genes reaction(-1). Reliable quantification of the target AOB DNA was obtained when the target AOB DNA comprised more than 0.1% of total AOB DNA in the sample. The application of the primer sets to samples taken from five sewage treatment systems showed that, in all systems, the majority of the AOB population was comprised of one sequence type of the N. oligotropha cluster (3.9+/-1.5x10(9)-1.7+/-0.5x10(10) cell l(-1)) and, in most systems, followed by members within the N. communis cluster (2.8+/-0.3x10(9)-1.0+/-0.1x10(10) cell l(-1)) or/and another sequence type of the N. oligotropha cluster (1.5+/-0.6x10(8)-5.5+/-0.5x10(8) cell l(-1)). N. europaea-N. mobilis cluster arose solely in small numbers (4.9+/-0.8x10(8) cell l(-1)) in o...Continue Reading

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Citations

Oct 6, 2010·Applied Microbiology and Biotechnology·Puntipar Sonthiphand, Tawan Limpiyakorn
Jan 12, 2013·Applied Microbiology and Biotechnology·Tawan LimpiyakornPuntipar Sonthiphand
Feb 5, 2014·Applied Microbiology and Biotechnology·Jingyin WangHongwei Rong
Oct 12, 2007·Biotechnology and Bioengineering·T S Radniecki, R L Ely
Jan 10, 2017·Water Science and Technology : a Journal of the International Association on Water Pollution Research·Takahiro WatariTakashi Yamaguchi
May 8, 2020·The Science of the Total Environment·Siqi LiMin Zheng

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