Anaerobic granular sludge and removal of ibuprofen in sanitary sewage using ethanol cossubstrate
Genomic DNA extraction from granular sludge samples was performed with FastDNA  SPIN Kit for Soil DNA Extraction (MP Biomedicals). The sequencing was performed by the company Genone of Rio de Janeiro-RJ (Brazil). The primer sets used for the 16S RNAr gene were 341F / 806R (V3V4 region) for the Bacteria Domain and U519F / 806R (V4 region) for the Archaea Domain.  Sequences analysis were performed by Uparse software (Uparse v7.0.1001?http://drive5.com/uparse/) (Edgar 2013). Sequences with ?97% similarity were assigned to the same OTUs. Representative sequence for each OTU was screened for further annotation. For each representative sequence, the GreenGene Database (http://greengenes.lbl.gov/cgi-bin/nph-index.cgi) (DeSantis et al. 2006) was used based on RDP3 classifier (Version 2.2, http://sourceforge.net/projects/rdp-classifier/) (Wang et al. 2007) algorithmto annotate taxonomic information. In order to study phylogenetic relationship of different OTUs, and the difference of the dominant species in different samples (groups), multiple sequence alignment were conducted using the MUSCLE software (Version 3.8.31?http://www.drive5.com/muscle/) (Edgar 2004). All this indices in our samples were calculated with QIIME (Version 1.7.0) and displayed with R software (Version 2.15.3).