Molecular Identification of Locally Isolated Bacteria from Soil in Thiqar Governorate and Ability to Produced Antibiotics

Abstract

AbstractThe objective of this study was to isolate and diagnose local bacterial isolates producedantimicrobials from some soil in Thiqar governorate and detecting the presence of 16SrRNA geneusing DNA Sequence technique. The study included collected 25 soil samples from different regionsof Thiqar Governorate, 13 bacterial isolates wasisolated depending on shapes and colors, RL1 andRL2.These isolates were identified by biochemical tests. They were identified as Bacillus isolatesand were also identified using Vitek 2. The technique was able to diagnose only 7 isolates. Theisolates were then tested to produce antimicrobial agents against 5 known bacterial pathogensKlebsiella pneumoniae, Proteus mirabilis, Pseudomonasaeruginosa, Staphylococcus aureus,Salmonella typhi. Most isolates showed a positive result against these pathogenic bacteria and thenisolated 9 isolates using 16SrRNA gene for PCR products. All isolates were positive for this gene.After treatment for each isolate using Mega 6 program, and obtained new isolate and get codeMF423696 after matching them with NCBI. And 5 isolates were selected for the purpose ofcompleting the study. The antibiotic was then extraction by centrifugation at 10.000 cycles / minutefor 15 minutes and then filtered using 0.22μm micrometer filter. It was used to inhibit the pathogenicbacteria. A set of specific reagents was carried out to identify the active substances found in theextract including detection of phenols, flavonoids, amines, and saponins. All isolates were positivefor phenols, flavonoids, amines, and saponins except RL2 that showed negative to this test.