Characterization of microbial communities in a pilot-scale constructed wetland using PLFA and PCR-DGGE analyses

Abstract

Phospholipid fatty acid (PLFA) analysis and 16S ribosomal DNA polymerase chain reaction amplification-denaturing gradient gel electrophoresis (PCR-DGGE) were used to determine microbial communities and predominant microbial populations in water samples collected from a pilot-scale constructed wetland system. This pilot-scale constructed wetland system consists of three types: subsurface-flow (SSF), surface-flow (SF) and a floating aquatic plant (FAP) system. Analysis of PLFA profiles indicated primarily eukaryotic organisms, including fungi, protozoa, and diatoms were observed in all three wetland systems. Biomarkers for Gram-negative bacteria were also detected in all samples analyzed while low proportions of biomarkers for Gram-positive bacteria were observed. Biomass content (total PFLA/sample) was highest in water samples collected from both SF and FAP system while highest metabolic activity was observed in FAP system. This is consistent with the observed highest metal removal rate in FAP system. Sequence analysis of the predominant PCR-DGGE DNA fragments showed 0.92 to 0.99 similarity indices to Beta-proteobacteria, Flavobacterium sp. GOBB3-206, Flexibacter-Cytophaga-Bacteroides group, and Gram-positive bacteria. Results suggest diverse microbial communities including microorganisms that may significantly contribute to biogeochemical elemental cycles.