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E.coli was reported as the most predominant uro-pathogen, which may be difficult to treat as they exhibit multidrug resistance and sometimes grow in biofilms. Because honey exhibits antimicrobial activities against a wide range of bacteria, this study was undertaken to study resistance pattern and biofilm formation among uropathogenic E.coli, and evaluate the use of Egyptian honey as an antibacterial agent. Uropathogenic E.coli strains were collected and susceptibility test by commercial antibiotic was performed. Extended-spectrum beta-lactamase (ESBL) detection was assessed by confirmatory double-disc assay.
Biofilm formation was detected by the Tissue Culture Plate method. Antibacterial activity of four types of Egyptian honey (citrus, camphor, marjoram, and black seed honey) was tested by agar well diffusion and micro-broth dilution methods. 17 uropathogenic E. coli strains were collected, at which high resistance rates toward ampicillin (100%) followed by cephalosporins such as ceftazidime (88%) and cefotaxime (77%) were detected. Imipenem was the most active antibiotic with susceptibility of 94%. 11 isolates (64.7%) were ESBL producers while only 6 isolates (35.
3%) were non-ESBL producers.
12 isolates (70.6%) were found to be biofilm formers at which 3 (17.7%) of them were categorized as moderate biofilm formers and 9 (52.9%) were categorized as weak biofilm formers. Only 5 isolates (29.4%) were non-biofilm formers. 50% citrus honey concentration was defined as MIC for 3 isolates, and a concentration of 12.5% was defined as MIC for 1 isolate. Our study showed high resistance rates of uropathogenic E.coli toward the used antibiotics, in addition to ESBL production and biofilm formation ability. Egyptian citrus honey at a concentration of 50% and 12.5% had the ability to inhibit 23.53% of E.coli isolates, acting as a good antibacterial agent against such resistance pathogenic strains of E.
Among the most common infections, urinary tract infections (UTIs) are commonly encountered diseases by clinicians in developing countries
Honey is the natural sweet substance from nectar or from the secretion of living parts of plants or excretions of plants, which honey bees collect, transform, and combine with specific substances of their own to ripen and mature 12. The use of honey as a drug for the treatment of disease dates back to 2100-2000 BC. For instance, pale honey was described by Aristotle (384-322 BC) as being ‘good for sore eyes and wounds’ 13. It was already found by many scientists that, honey has the most powerful inhibitory effect with regard to sixty species of bacteria. Among them, many bacteria are resistant to antibiotics but not towards honey. Because it is non-toxic and will not produce any adverse effects 14. In the context of this, the present study was undertaken with the aim to study resistance pattern and biofilm formation among uropathogenic E.coli, and evaluate the use of Egyptian honey as an antibacterial agent.
17 uropathogenic E.coli isolates were kindly collected from Kasr Alaini Hospital, Egypt. All isolates were collected on Cled and MacConkey agar. Isolates were defined as E.coli according to routine biochemical testing 15. After confirmation of isolates identification, they were collected on glycerol broth and stored at -80 freezer until been used.
Antimicrobial susceptibility of the E. coli isolates was determined by the disk diffusion as recommended by CLSI 16. The commercial antibiotics tested were imipenem (10Вµg), cefotaxime (30Вµg), cefoxitin (30Вµg), gentamicin (10Вµg), amikacin (30Вµg), ciprofloxacin (5Вµg), levofloxacin (5Вµg), ceftazidime (30 Ојg), and ampicillin (10Вµg). Results were interpreted according to CLSI criteria.
ESBL confirmatory test was done to all isolates using combination disk method (Bio-Rad, France) as recommended by CLSI17 and include cefotaxime (30 Ојg), cefotaxime/ clavulanic acid (30/10 Ојg), ceftazidime (30 Ојg), and ceftazidime/ clavulanic acid (30/10 Ојg). Klebsiella pneumoniae standard strain ATCC 700603 was used as a positive control.
Evaluation of biofilm formation by E.coli was performed according to the method described by Mohamed, et al. 18 using Tissue Culture Plate (TCP) method with minor modifications. Wells originally containing uninoculated media were considered as negative control. The optical density (O.D) was measured at 630 nm using STAT FAX 2100 microplate reader. The cut-off value (ODc) is defined as three standard deviations (SD) above the mean OD of the negative control, that is, sample’s ODc = average OD of negative control + (3*SD of negative control). After comparing the O.D of biofilm to the control and according to the readings, the isolates were classified as follows: O.D в‰¤ O.Dc no biofilm producer, O.Dc
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