Bioremediation of Organic Contamination Using Molecularly Identified Bacillus Subtilis Bacteria

Abstract

Soil bioremediation primarily depends on the enzymatic activity of living microorganisms, to facilitate the degradation of contaminants or their conversion into less dangerous forms. Ten soil samples were collected from different locations in the city of Sharqat and the Rumana area of Qayyarah district. The Physical (concentration and Electrical Conductivity) and chemical tests (pH) conducted on soil samples. The B. subtilis were cultured and identified by Gram stain and VITEK2 compact system, also DNA extraction and Genome sequencing of 16S RNA ribosome gene of bacteria was carried out to complete the diagnosis of B. subtilis. Bacillus subtilis is a characteristic bacterium that is rod-shaped. When cultivated on standard nutritional agar, the shape of this bacterium's circular colony is rough, opaque, fuzzy white or slightly yellow, exhibiting jagged edges. Sequencing revealed that most of the isolated strains were B. subtilis. The result of PCR showed that U1 gene has found in 100% of the B. subtilis isolates. BLAST results show 98% nucleotide sequence similarity and similarity for the 16RNA ribosomal gene of B. subtilis with no gaps. The sequence for 16RNA ribosomal gene was sub-mitted to NCBI, Gene Bank with accession No. KY962959.1. The alignment was done through Clustal Omega that recognized partial variations on some bases. The concentration of spontaneous wastes was decreased with bioremediation (3.864) for controlled temp. and (3.21) with un controlled temp. the Electrical Conductivity also decreased (3647) for controlled temp. and (2265) with uncontrolled temp. The pH increase in controlled temp. (8.699) and uncontrolled temp. (8.23). B.subtilis has a large efficiency in bioremediation process which had a benefit in ecosystem. It has a higheffect in uncontrolled environmental condition(temperature) which lead to decrease the concentration ofSpontaneous wastes and decease the electrical conductivity and increase the pH to more alkaline pH.