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Abstract

DETERMINATION OF EXTENDED SPECTRUM BETA LACTMASE ACTIVITY IN UROPATHOGENS IN DIABETIC PATIENTS BY DOUBLE DISK SYNERGY METHOD (DDS)

Betty Daniel and Mehvish Saleem*

Abstract

Background: Urinary tract Infection (UTI) is the most common infection observed in patients with diabetes related to the poor Immunological response and glycemic control. Extended spectrum Beta lactamases (ESBLs) is an excellent mechanism of bacterial resistance adopted especially by gram negative bacteria responsible for UTI. Aim This study determines the prevalence of ESBL activity among uropathogens conferring resistance to antibacterial drugs commonly used in the treatment of UTI. Material and Methods: A total of 240 isolates of E.coli, Pseudomonas aeruginosa, and Staphyloccus aureus were isolated from urine samples collected from Bangalore Hospital between the period of 2010-2011. Mid-stream urine or the clean catch urine sample was collected. The urine was streaked on the surface of Hichrome UTI Agar (Himedia Labs India pvt ltd) using a sterile inoculation loop. The plates were incubated at 37c. Colonies were identified on the basis of the color and standard biochemical methods. Staphylococcus aureus was identified on blood agar for beta-hemolysis and coagulase test. Disc diffusion method was used to determine the antimicrobial susceptibility. Determination of ESBL production was phenotypically confirmed by the Double disk synergy method (DDS) following CLSI recommendations. first, a suspension was prepared for each pure bacterial isolate according to the 0.5 McFarland turbidity standard and cultured on Mueller–Hinton agar. The test was performed on agar with a 30-μg disk of cefotaxime (and/or ceftriaxone and/or ceftazidime and a disk of amoxicillin–clavulanate (containing 10 μg of clavulanate) positioned at a distance of 30 mm (centre to centre), i.e., at the distance provided by several types of disk-dispenser. The test was considered as positive when a decreased susceptibility to cefotaxime was observed with a clear-cut enhancement zone of inhibition of cefotaxime in front of the clavulanate-containing disk, showing a characteristic shape-zone referred to as „champagne-cork‟ or „keyhole‟. Results: Among 240 isolates tested isolates of E.coli 145(60%) were resistant to Ampicillin, Nalidixic acid, ceftriaxone, ceftazidime, aztronam, Norfloxacin, ofloxacin and Cotrimoxazole. 95 isolates (39%) were sensitive to Fosfomycin, Gentamicin, Imipenem, ciprofloxacin, ofloxacin, levofloxacin and Nitrofurantoin. 180 isolates of Pseudomonas aeruginosa (75%) were sensitive to Amikacin, Tobramycin, Ciprofloxacin, levofloxacin and Fosfomycin and 60 isolates(25%) were resistant to Ceftazidime, Cotrimoxazole, Polymixin B, Norfloxacin and Ofloxacin. Among the Staphylococcus aureus isolates 168 isolates (78%) were resistant to amoxiclav, cotrimoxazole, ciprofloxacin, amikacin, norfloxacin, and nitrofurantoin. 72(30%) isolates were sensitive to Methicillin, Gentamicin, Amikacin, Fosfomycin, ciprofloxacin and vancomycin. The percentage of ESBL producing organisms was E.coli =26%, Pseudomonas aeruginosa =30% and Staphylococcus aureus= 25%.

Keywords: Diabetes mellitus (DM), Urinary tract Infection (UTI), Extended spectrum Beta Lactamases (ESBLs), Uropathogens, Antibacterial resistance.


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