Preparation of hybrid Nano-ciprofloxacin and determination its inhibition activity against E. coli isolated from urinary tract infection patients in holy Karbala city

Background: Urinary tract infections are the most common infections after upper respiratory tract infections E. coli caused 20% of urinary tract infection and it was responsible for 1/3 of the infections. Nanoparticles (NPs) in general have many properties that are different from those of traditionally used materials. They have dimensions typically below 100 nm, which allows them to reach specific sites inside the body and even to be permeable to tissues and cells. Therefore, they can deliver the drugs in active forms at sites that conventional drugs may not reach by themselves and thus minimize the undesirable side effects. Layered double hydroxides (LDHs) as one of the inorganic delivery nanoparticles have recently been receiving considerable attentions. Ciprofloxacin is a broad-spectrum antibiotic of the fluoroquinolone class. It is active against both Gram-positive and Gram-negative bacteria and it is recognized as one of the most effective antibiotics of the quinolone drug class and has been used for the treatment of urinary tract infections. Aims and objectives: The aims of present study were to prepare hybrid nanoan-ciprofloxacin and characterize by spectroscopic methods, and determine its inhibitory activity against the E. coli isolates caused the urinary tract infection in this study. Materials and Methods: In this study urine samples from 75 patients clinically diagnosed with urinary tract infection were collected into sterile screw capped container and used for culture, Gram staining, API E20, antibiotic sensitivity test and singleplex PCR methods. PCR was performed with primer pair targeted to the PhoA gene of E. coli. Hybrid nano-antibiotic represented by the Mg/Al-Ciprofloxacin-LDH was prepared by using the direct ion exchange method between the free ciprofloxacin antibiotic and Mg/Al-LDH carrier, and then the hybrid nano-ciprofloxacin was characterized by spectroscopic methods include FTIR, XRD and AFM. Results: Out of 75 samples, 27(36%) samples have been detected for E. coli depending on the morphologic and biochemical characteristics of this bacterium on the nutrient agar, MacConky agar, blood base agar and EMB agar, as well as Gram stain and API E20. The antibiotic sensitivity test was showed different sensitivity patterns of E. coli isolates against the different types of antibiotics varied from marked sensitive pattern against imipenem and meropenem, to moderate sensitive pattern against ciprofloxacin, norfloxacin, levofloxacin, nitrofurantoin and amikacin, in addition, to the marked resistance pattern toward ampicillin, ticarcillin, amoxicillin-clavanic acid and piperacillin. The singlplex PCR positive samples were identified by presence of ~903bp amplicon of the PhoA gene. The results of FTIR, XRD and AFM methods of nano-ciprofloxacin characterization indicted that this prepared nano-antibiotic was hybrid nano-ciprofloxacin (Mg/Al-Ciprofloxacin-LDH). The results of inhibition activity of both free ciprofloxacin and hybrid nano-ciprofloxacin showed that these two forms of ciprofloxacin revealed highest inhibition activity against two E. coli isolates (IS8 and IS10) with diameter of inhibition zone 40 and 42 mm, respectively. Whereas the E. coli isolates IS1and IS2 were the least affected isolates by the inhibition activity of these two forms of cirofloxacin with diameter of inhibition zone 38 and 35 mm, respectively. Conclusion: Using of hybrid nano-antibiotics resulting from loaded of different antibiotics on LDHs carrier could be represented a promising future tool to treat and control urinary tract infection and other different bacterial infections.