Abstract
Aim: To determine the Plasmid DNA profile of the multidrug resistant strains of Pseudomonas aeruginosa in the clinical isolates. Materials and Methods: Of the 150 clinical samples (Ear swab, Urine, Wound swab, Sputa and Semen) received at Lahor Research Laboratory and Medical center in Benin City, between January 2010 and December 2012, 36 (24%) yielded significant growth of P. aeruginosa. Samples were cultured on MacConkey and Blood agar. Clinical isolates were identified using standard method. Antibiotics susceptibility test employing agar disc diffusion method was used. Clinical isolates were subjected to Plasmid DNA profiling and curing test was carried out at Lahor Molecular Laboratory. This was followed by a post plasmid curing susceptibility test. Agarose gel electrophoresis was carried out to separate the Plasmid DNA using standard method. Bands were visualized using UV illuminator. Results: Wound swabs had the highest numbers of clinical isolates of P. aeruginosa (55.6%) followed by Urine, Semen, Sputa and Ear swab (19.4%, 11.0%, 8.3%, and 5.6%) respectively. Before the isolates were cured of their plasmid, 39% of the P. aeruginosa strains were found to be resistant to Ciprofloxacin (CPX), 47%, Ofloxacin (OFX), 44% Pefloxacin (PEF) and 56% Sparfloxacin (SPX). After plasmid curing, the new antibiogram of the isolates showed that some clinical isolates that hitherto were resistant to a given Fluoroquinolone became susceptible, 36% to CPX, 12% to OFX, 12.5% to PEF and 15% to SPX. Agarose gel electrophoresis carried out on the Plasmid DNA revealed that there was detectable Plasmid DNA in 13.9% of the clinical isolates analyzed. Conclusion: There is an alarming increase of clinical infections caused by multidrug resistant strains of P. aeruginosa.13.9% of the multidrug resistance strains of P. aeruginosa in Benin City were Plasmid mediated. Treatment should be based on current Laboratory Susceptibility Test results of the isolates.
Highlights
Pseudomonas aeruginosa is a Gram-negative aerobic, coccobacillus bacterium with unipolar motility
Plate 1: Shows the Results of the Plasmids DNA profiling analyzed in a 0.8% Agarose gel Electrophoresis
Our study revealed that wound swab had the highest number of clinical isolates of P. aeruginosa (55.6%) followed by urine sample (19.4%), Semen (11%) and Sputa sample (8.3%)
Summary
Pseudomonas aeruginosa is a Gram-negative aerobic, coccobacillus bacterium with unipolar motility. In mixed cultures, it can be isolated as clear colonies on MacConkey agar (as it does not ferment lactose) which will test positive for oxidase [1]. It can be isolated as clear colonies on MacConkey agar (as it does not ferment lactose) which will test positive for oxidase [1] It is often preliminarily identified by its pearlescent appearance and grape like odor invitro. Definitive clinical identification of P. aeruginosa often includes identifying the production of pyocyanin and fluorescein as well as its ability to grow at 42 ̊C. P. aeruginosa is capable of growth in diesel and jet fuel, where it is known as a hydrocarbon-using microorganism OJMM.
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