Paper Title
Sequencing and Molecular Characterization of Plasmid-Mediated Antibiotic-Resistant Bacteria Encoding Shv and Tem Genes from some Cooked Street Food Sold in Osun, A South Western Nigeria State.
Abstract
sold in Osun, Nigeria were studied. 75 bacteria were isolated and identified using conventional techniques. Resistance patterns of the obtained isolates were studied using; Ceftazidime (30μg), cefruroxime (30μg), gentamicin (10μg), ciprofloxacin (5μg), ofloxacin (5μg), cefixime (5μg), amoxicillin (30μg), nitrofurantoin (300μg), and augmentin (30μg).Curing of plasmids and DNA extraction procedures were performed on bacterial isolates resistant to four or more antibiotics.The isolates with several antibiotic resistance genes on their plasmid were amplified (PCR methods) and identified using the 16S rRNA gene and NCBI blast. Using Illumina methods, one full plasmid DNA of an identified bacteria was sequenced and analyzed using snapgene and the geneious prime bioinformatics tool. Antibiotic resistance was observed to ceftazidime (77%), augmentin (88%) and cefuroxime (73%), while sensitivity to ciprofloxacin and nitrofurantoin was at 93% and 91% respectively. Twelve of the bacteria identified carried multi-antibiotic resistance, including the presence of SHV-1, SHV-2, TEM-1, TEM-2, and OXA2 in the bacteria. Using the KmerRes finder database, bioinformatics analysis of Alcaligenesfaecalis complete plasmid sequence (identified using 16S rRNA and NCBI database) through assembly, annotation, and map identified some enzymes as well as other resistance genes (like fosB, tet, Isa(B), aph2) carried by the bacteria.
Drug treatment failures have increased as new resistance features in food-borne bacteria have been discovered. However, combining sequencing techniques with bioinformatics tools has improved our knowledge of the full tailored information contained in such bacterium genomes, allowing us to find new effective treatment options.
Keywords - Plasmid DNA, 16S RRNA, resistance genes, Bioinformatics tools