Molecular characterization and plasmid profiling of bacteria isolates for bioremediation, from Naraguta leather works wastewater of Jos, Plateau State, Nigeria

Abstract

Environmental contamination is widely a global issue due to increased industrialisation and other human activities. It is therefore important to identify bacteria in that environment with contaminant degrading abilities, which could serve in bioremediations. Therefore, this study aimed to carry out isolation and plasmid profile of bacteria from contaminated water of leather works facilities. The effluent was collected and cultured on Nutrient Agar and the pure colonies were further transferred to cetrimide agar then to nutrient broth. The isolates were characterised microbiologically. The isolates were also subjected to biochemical analysis including Oxidase, Urease, Catalase, indole and citrate tests. The extraction of DNA from the pure isolates was carried out using the ZymoBead^TM DNA Extraction Kit. Polymerase chain reaction (PCR) was carried out to amplify the 16S rRNA gene of the isolates. Agarose gel electrophoresis was carried out to visualise the output of the PCR reaction using 1% agarose.  Unpurified PCR products were sent for sequencing. The sequences generated were further analysed using bioinformatics tools.  Plasmid isolation was carried out on the isolates using Monarch® Plasmid DNA Miniprep Kit and it was then run on gel electrophoresis to visualise the output. The bioinformatics results showed the organisms Pseudomonas aeruginosa strain S1C1 (100%), Pseudomonas aeruginosa SSK113J146 (96%), Pseudomonas aeruginosa strain PA-W (95%), Pseudomonas aeruginosa strain PWA 16SR (95%) are the closer organism to A3a, while Stenotrophomonas maltophila strain M2 (100%), and Stenotrophomonas sp. NGB23 (100%), Stenotrophomonas maltophila ESG6 (92%), Stenotrophomonas maltophila strain ISSDS-759 (92%), Unculture bacterium clone ncd1606d04c1 (92%), Stenotrophomonas maltophila strain BHWSL2 (100%) are closer to C1a. Query cover was used for close identity. Plasmid profiling showed that the isolates have one plasmid each with an approximate size of 4kb. In conclusion, the isolates as compared with the database need further check.