Bacterial community analysis by PCR-DGGE and 454-pyrosequencing of horizontal subsurface flow constructed wetlands with front aeration

Abstract

Horizontal subsurface flow constructed wetlands (HSSF CWs) with and without redox manipulation by front aeration were operated to treat mechanically pretreated wastewater from a nearby wastewater treatment plant. Polymerase chain reaction-denaturing gradient gel electrophoresis and 454-pyrosequencing were used to characterize the shifts in bacterial community diversity and composition in response to front aeration in the HSSF CWs. Both techniques revealed similar bacterial diversity between the HSSF CWs with (ACW) and without front aeration (NACW). Differences in microbial functional groups between the ACW and the NACW substrate samples were identified with 454-pyrosequencing. Nitrite-oxidizing bacteria (Nitrospira) and ammonia-oxidizing bacteria (Nitrosomonas) had much higher abundances in the ACW, whereas more sequences related to sulfate-reducing bacteria and anaerobic sulfur-oxidizing bacteria (genera Sulfuricella, Sulfuritalea, and Sulfuricurvum) were detected in the NACW. Removal efficiencies for NH₄(+)-N, PO₄(3-)-P and chemical oxygen demand in the ACW were 48.7 ± 15.5, 70.2 ± 13.5, and 82.0 ± 6.4%, respectively, whereas the removal efficiencies for these parameters in the NACW were 10.3 ± 14.0, 53.1 ± 18.9, and 68.8 ± 10.7%, respectively. In the ACW, the stimulation of nitrification via front aeration supplied more NO₂(-)-N and NO₃(-)-N to the subsequent denitrification process than in the NACW, resulting in higher total inorganic nitrogen removal efficiency. The differences in treatment efficiencies between the ACW and the NACW could be partially explained by the different bacterial community compositions in the two CWs.

Characteristics of Bacterial Communities in Cyanobacteria-Blooming Aquaculture Wastewater Influenced by the Phytoremediation with Water Hyacinth

“…After cyanobacteria-blooming aquaculture wastewater was treated by water hyacinth for 30 days, Proteobacteria was found to be the predominant phylum in total bacteria of water with a relative abundance of 61.27 ± 3.97%. As reported, Proteobacteria is one of the most common bacterial phylum in aquaculture ponds [23] and is also an important microorganism involved in global carbon and nitrogen and sulfur cycle [24]. Its relative abundance in constructed wetland is always particularly high and up to 50-61% [25].…”

Microbial abundance and community in subsurface flow constructed wetland microcosms: role of plant presence

“…Numerous studies have realized the pyrosequencing of 16S rRNA V3 hypervariable regions of the bacteria in CWs for quantifying bacterial populations and characterizing microbial community diversity using PCR-DGGE technique or 454 sequencing platform with wastewater treatment (Zhong et al 2014;Faulwetter et al 2013;Dong and Reddy 2010). In the current study, the 16S rDNA hypervariable V3 region was amplified through PCR with the universal bacterial primers 341f (Watanabe et al 2001) and −518r (Muyzer et al 1993).…”

Influence of substrate type on microbial community structure in vertical-flow constructed wetland treating polluted river water

“…High-throughput sequencing technology is a highly efficient tool for identifying the profile of complicated microbial community (Liao et al 2015;Wang et al 2015a, b;Yang et al 2015), and can provide a new opportunity to resolve the microbial assemblage of CW biofilm and systematically investigate its links with operational and environmental conditions (Ansola et al 2014;Arroyo et al 2015;Zhong et al 2015). Due to its relatively low costs and great throughput, Illumina sequencing has been applied to profile microbial communities in various natural and man-made ecosystems Shi et al 2014;Sun et al 2015;Wu et al 2015;Yang et al 2014;Zhang et al 2015).…”