Use of quantitative real-time PCR to monitor population dynamics of ammonia-oxidizing bacteria in batch process |
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Authors: | Juntaek Lim Seungyong Lee Seokhwan Hwang |
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Institution: | (1) School of Environmental Science and Engineering, Pohang University of Science and Technology, Pohang, Gyungbuk, 790-784, South Korea |
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Abstract: | A quantitative real-time PCR (QPCR) assay with the TaqMan system was used to quantify 16S rRNA genes of β-proteobacterial
ammonia-oxidizing bacteria (AOB) in a batch nitrification bioreactor. Five different sets of primers, together with a TaqMan
probe, were used to quantify the 16S rRNA genes of β-proteobacterial AOB belonging to the Nitrosomonas europaea, Nitrosococcus mobilis, Nitrosomonas nitrosa, and Nitrosomonas cryotolerans clusters, and the genus Nitrosospira. We also used PCR followed by denaturing gradient gel electrophoresis (DGGE), cloning, and sequencing of their 16S rRNA genes
to identify the AOB species. Seed sludge from an industrial wastewater treatment process controlling high-strength nitrogen
wastewater (500 mg/L NH4
+–N) was used as the inoculum for subsequent batch experiment. The Nitrosomonas nitrosa cluster was the predominant AOB (2.3 × 105 copies/mL) in the start-up period of the batch experiment. However, from the exponential growth period, the Nitrosomonas europaea cluster was the most abundant AOB, and its 16S rRNA gene copy number increased to 8.9 × 106 copies/mL. The competitive dominance between the two AOB clusters is consistent with observed differences in ammonia tolerance
and substrate affinity. Analysis of the DGGE results indicated the presence of Nitrosomonas europaea ATCC19718 and Nitrosomonas nitrosa Nm90, consistent with the QPCR results. |
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Keywords: | Ammonia-oxidizing bacteria Industrial wastewater Nitrosomonas Population dynamics Quantitative real-time PCR |
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