Isolation and molecular identification of Clostridium bifermentans from anaerobic lagoons of wastewater treatment system

Abstract

 Introduction: Clostridiumbifermentans is a non pathogenbacteria and first isolated by Tissier and Martelli in 1992. This bacteria with a view to taxonomy has a close relationship with the Clostridiumsourdilli and is able to produce a wide range of metabolites such as acetic acid, lactic acid, formic acid, ethanol, butanol, acetone, carbon dioxide, nitrogen and specially hydrogen as clean source energy. The aim of this study was to isolate and molecular identify Clostridium bifermentans. Materials and methods: With aim to isolation of Clostridium bifermentans, the anaerobic wastewater samples from Mahdishahr sewage treatment laggons cultured on asynthetic complex medium and incubated anaerobically at room temperature. Chromosomal DNA isolated from grown bacteria and the 16s rRNA gene amplified with PCR. The PCR product was sequenced and purified. Results: Isolated bacteria from a macroscopic view on the complex media has white colonies and from a microscopic view was rod-shaped Gram-positive and had spore. of the 16s rRNA gene sequencing, isolated bacteria belong to the species Clostridium bifermentans which is called IRI2 strain and were recorded in gene bank with acception number KC525132.  Discussion and conclusion: In this study a strain of Clostridium bifermentans was separated and identified with 16S rRNA sequencing method. With respect to the ability of this bacteria to produce various industrial products, this bacteria is to be used especially for production of hydrogen as biogas. 

Keywords

• Clostridium bifermentans
• Wastewater
• Identification
• 16SrRNA