Curing Analysis of Drug Resistance Plasmid in Proteus Mirabilis

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Author: Lawin Ahmed Omer, Zirak Faqe A. Abdulrahman,
Department of Biology, College of Education, University of Salahaddin – Hawler
Author: Rastee Hasan Saeed
Department of Biology, College of Education, University of Salahaddin – Hawler , Cihan University-Erbil

DOI: http://dx.doi.org/10.24086/cuesj.si.2017.n2a5

Abstract

Forty five isolates of Proteus mirabilis were identified among 600 samples taken from patients with urinary tract infection from different hospitals in Erbil City.  Isolates were identified from urine sample by using cultural, morphological and biochemical characteristics and confirmed confirmed by VITEK 2 systems. Antibiotic sensitivity testing was done for all isolates by using fifteen antibiotic types which included Amikacin (AK), Ampicillin (AMP), Aztreonam (AT), Chloramphenicol (C), Cephalothin (CEP), Ciprofloxacin (CIP), Ceftriaxone (CTR), Cefotaxime (CTX) Fosfomycin (FOX), Gentamicin (GEN), Imipenem (IMP), Nalidixic acid (NA), Norfloxacin (NOR), Piperacilin (PI) and Tobramycin (TOB). IMP was the most effective antibiotic against isolated of P. mirabilis. The resistance rate of the isolates toward these antibiotics were 8.8% for CIP, 11% for AT, 11% for FOX, 15.5% for NOR, 17.7% for TOB, 20% for AK, 22% for PI, 26% for CTR, 28.8% for CTX, 33% for NA, 35.5% for GEN, 44% for C, and 62% for AMP respectively. To control the antibiotic resistance of the tested P. mirabilis isolates, curing of plasmid DNA was conducted using tetracycline and elevated temperature at 46 ^oC, two most resistance isolates were chosen for this purpose P32, and P40, then treated with different concentrations of tetracycline (0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4) µg/ml separately. The MIC for tetracycline was 2.5µg/ml and SMIC 2µg/ml was used as a curing agent. The genes encoded resistance to ampicillin, chloramphenicol, cephalothin, ceftriaxone, cefotaxime, gentamicin and tobramycin were cured from P32 and the percentage of curing was (46.6%), while amikacin, chloramphenicol, ciprofloxacin and gentamicin resistance genes were cured from P40 isolate and the curing percent was (26.6%).The results confirmed by conducting gel electrophoresis and revealed that tetracycline removed plasmid of P32, while it had no effect on plasmid of P40. On the other hand, elevated temperature used also as curing agent and the results revealed that resistance to amikacin, ampicillin, chloramphenicol, cephalothin, ceftriaxone, gentamicin and tobramycin genes were cured from P32 and the curing percent was (46.6%), while genes encoding amikacin, ampicillin, chloramphenicol, ciprofloxacin, ceftriaxone, cefotaxime, fosfomycin, gentamicin, nalidixic acid, norfloxacin and tobramycin were cured from P40 isolate with the percentages of (73.3%) after incubating the isolates at 46 ^o C. It is clear that elevated temperature is the most efficient method than tetracycline. The results confirmed by conducting gel electrophoresis and showed that both tested isolates lost their plasmids after incubation at 46 ^o C.

Key words: Proteus mirabilis, antibiotic resistance, and plasmid curing

 

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