Identification and potential application of bacterial population in biogranule for the treatment of textile wastewater

Abstract

Identification of bacterial population from biogranules for remediating industrial wastewater has been carried out. In this study, molecular method of metagenomic was applied utilizing the 16S rRNA gene fragment to determine the diversity of bacterial population. The genomic DNA was directly extracted from biogranules. The 16S rRNA gene was amplified by PCR and universal primers. Transformation was done into JM 109 competent cell by heat-shock method for selecting the positive clones and gaining high number of cloned vectors. In order to recover the individual positive clones, the plasmid was extracted, for confirming the cloning from plasmid gel electrophoresis method was conducted. Then, reconfirmation by restriction enzyme was performed by EcoR1, after cutting the vector and the gene was separated at the band of 3000 bp for vector and 1500 bp for gene. Finally, 12 positive colonies were observed from extraction of plasmid. The positive colonies were sent for commercial sequencing, and the sequences obtained were then analyzed using bioinformatics tools in order to identify the bacterial genus and species as well as to locate them in the phylogenetic tree. Three main bacterial phylogroups including: Firmicutes, Spirochaetes, and proteobacteria as well as different species of bacterial were finally identified. In the second phase of this study, assessing of the biogranules performance in textile wastewater was accomplished. The parameters such as Ammonical-Nitrogen, Colour (ADMI), COD removal and pH have been analyzed.