Microbial processes play a vital important role in the removal of contaminants in constructed wetland (CW). However, the microbial physiology and community structure can be influenced by environmental conditions. In this study, four pilot-scale integrated vertical-flow constructed wetlands (IVCWs) were employed to treat domestic and nitrified wastewaters. The microbial properties, along with their response to wastewater quality characteristics and seasonal variation, were determined. The results showed higher Shannon-Weiner diversity (H) and evenness (E) index of fatty acids (FAs), and relative abundances of signature FAs in down-flow cells and in the systems fed with domestic wastewater (DW). The relative abundances of fungi and gram-negative and aerobic bacteria were greater in up-flow cells. The dominant anaerobic bacteria found in most cells might be accounted for the prevailing anaerobic environment within the wetland beds, which could mean that the system fed with nitrified wastewater (NW) should perform better in nitrogen removal. The redundancy analysis (RDA) showed that pollutant concentrations, especially organic matter, influence the FA compositions greatly, and the most significant difference of microbial community structures was detected in down-flow cells fed with DW and up-flow ones with NW. The branched FAs, which could be used to represent anaerobic bacteria, were observed in down-flow cells treating DW and had a significant positive correlation with chemical oxygen demand (COD) concentration, probably suggesting the important role of anaerobic bacteria in organic matter degradation in the IVCWs. Seasonal variation, however, did not greatly influence the microbial community structure in the IVCWs.