Molecular Typing of 16S rRNA of Azadirachta indica and Ocimum gratissimum Resistant Pseudomonas spp. from Sabe in Oke-Ogun, Nigeria

Abstract

The aim of this study was to present the molecular typing of bacteria which were resistant to extract of the leaves of Ocimum gratissimum and Azadirachta indica and to investigate whether there was any relationship between non-sensitivity to antibiotics and the extract of plants of four isolates: Salmonella typhii, Escherichia coli, Pseudomonas aeruginosa and Pseudomonas monteilii. Four solvents used for the extraction of the active ingredients of the plants were Ethanol, N-Hexane, Water, Ethanol and Water combined in five different concentrations. The four microorganisms listed above were isolated from raw milk, Borehole water and soil. The isolates were variously subjected to conventional morphological, biochemical tests and subsequently molecular screening (Polymerase Chain Reaction (PCR) and Sanger sequencing method). Pseudomonas aeruginosa (KF530797) and Pseudomonas monteilii (KJ676707) identified were subjected to BLAST using pairwise alignment technique from NCBI database. The two plant extracts had earlier been documented as resistant to Escherichia coli and Salmonella spp. which were earlier isolated from milk and water, respectively. There was no zone of inhibition as observed in all concentrations of plant extract considered except a noticeable clear zone of inhibition against synthetic antibiotics in the order of Ofloxacin and Gentamycin. Although suspected gene markers from the two-soil isolate were suspected to be amplified around 1400 bp. The 16S rRNA sequences of Pseudomonas aeruginosa (94.43%) was somewhat more variable than Pseudomonas monteilii (99.93%) indicating a pairwise alignment of Pseudomonas monteilii to be more conserved than P. aeruginosa.