Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR

doi:10.1007/s11783-016-0846-x

Front. Environ. Sci. Eng.

2016, Vol. 10

Issue (4) : 7 https://doi.org/10.1007/s11783-016-0846-x

RESEARCH ARTICLE

Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR

Junqin PANG¹,Masami MATSUDA¹,Masashi KURODA¹,Daisuke INOUE¹,Kazunari SEI¹,Kei NISHIDA²,Michihiko IKE^1,^*(

)

¹. Division of Sustainable Energy and Environmental Engineering, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
². International Research Centre for River Basin Environment, University of Yamanashi, 4-3-11 Takeda, Kofu, Yamanashi 400-8511, Japan

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Abstract

Nitrogen-cycling microbial communities in municipal WWTPs were characterized.

Numbers of amoA, nirK and nirS genes were quantified by MPN-PCR.

Diversity of whole nitrogen-cycling communities was analyzed with DNA microarray.

CAS process retained diverse nitrogen cycling populations.

Specific, limited populations may be dominated in nitrogen removal processes.

To improve nitrogen removal performance of wastewater treatment plants (WWTPs), it is essential to understand the behavior of nitrogen cycling communities, which comprise various microorganisms. This study characterized the quantity and diversity of nitrogen cycling genes in various processes of municipal WWTPs by employing two molecular-based methods:most probable number-polymerase chain reaction (MPN-PCR) and DNA microarray. MPN-PCR analysis revealed that gene quantities were not statistically different among processes, suggesting that conventional activated sludge processes (CAS) are similar to nitrogen removal processes in their ability to retain an adequate population of nitrogen cycling microorganisms. Furthermore, most processes in the WWTPs that were researched shared a pattern:the nirS and the bacterial amoA genes were more abundant than the nirK and archaeal amoA genes, respectively. DNA microarray analysis revealed that several kinds of nitrification and denitrification genes were detected in both CAS and anaerobic-oxic processes (AO), whereas limited genes were detected in nitrogen removal processes. Results of this study suggest that CAS maintains a diverse community of nitrogen cycling microorganisms; moreover, the microbial communities in nitrogen removal processes may be specific.

Keywords DNA microarray analysis Nitrogen cycling functional genes Most probable number-polymerase chain reaction (MPN-PCR) Wastewater treatment plants (WWTPs)

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Corresponding Author(s): Michihiko IKE

Online First Date: 17 May 2016 Issue Date: 03 June 2016

Cite this article:

Junqin PANG,Masami MATSUDA,Masashi KURODA, et al. Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR[J]. Front. Environ. Sci. Eng., 2016, 10(4): 7.

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https://academic.hep.com.cn/fese/EN/10.1007/s11783-016-0846-x
https://academic.hep.com.cn/fese/EN/Y2016/V10/I4/7

Fig.1 Nitrogen cycle and its relevant functional genes

Tab.1 Water quality, activated sludge characteristics, and wastewater treatment performance of different WWTPs

Tab.2 Correlation coefficients between abundance of nitrogen-cycling functional genes and water quality, activated sludge characteristics, and wastewater treatment performance of processes.

Fig.2 MPN-PCR analysis to determine the abundance of eubacterial 16S rRNA and nitrogen cycling functional gene copies (bacterial amoA, archaeal amoA, nirK, and nirS) in activated sludge samples. (a) Quantities of gene copies in the samples. (b) Proportions of functional genes to eubacterial 16S rRNA gene. Error bar shows a 95% confidence interval

Fig.3 Quantity of gene probes detected in A-CAS (a), A-A2O (b), A-SFM (c), B-CAS (d), C-AO (e), D-CAS (f), and D-FCN (g) using DNA microarray analyses

Fig.4 PCA of nitrogen cycling functional gene composition obtained in microarray analysis in treatment processes

(a) Scatter diagram based on amo composition; (b) Scatter diagram based on the composition of denitrification genes

Fig.5 Percentage of positive probes for functional genes in nitrogen cycling in A-CAS, B-CAS, and D-CAS

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