1 Department of Biology, Science College, Mustansiriyah University, Baghdad, Iraq
2 Department of Microbiology, College of Health Sciences, Hawler Medical University
3 Department of Soil and Water Resources, Agriculture College, Kirkuk University
Background: The synthesis of metal nanoparticles using microorganisms has received great interest due to their optical, chemical, photoelectrical and electronic properties. The microorganisms are used as possible ―nanofactories‖ for development of clean, nontoxic and environmentally friendly methods for producing nanoparticles, the Nanoparticle are synthesized using various biosources such as bacteria, fungi, yeast, plant extract Objectives: The present study focuses on the biosynthesis of titanium nanoparticles using Iraqi Lactobacillus crispatus isolate and their inhibitory effect against multi drug resistance bacteria isolated from patient with Recurrent UTI and against their ESBL and Metallo beta lactamase. Materiales and Methods: Lactobacillus crispatuswas isolated from vagina of Iraqi healthy women then identified throughoutcultural, microscopical and biochemical test. Fifty samples of urine were collected from women suffering from recurrent UTI who had certain clinical symptoms; these samples were collected from Rizgary Teaching Hospital and Baghdad Teaching Hospital from August to September, 2013 with ages ranging from 18-60 years. The urine specimens were inoculated on both blood and MacConkeys agar plates. The isolates were identified throughout cultural, microscopical, biochemical test and Vitek 2 system. Isolates were tested against 14 antibiotics by using Kirby-Bauer disc diffusion and detection of Extend spectrum β- lactamases by double disc diffusion synergy method and detection of Metallo-β lactamase by disc potentiating test and Samples of synthesized nanoparticles were characterized after 72 hours of incubation. The formation of metal oxide TiO2 nanoparticles was confirmed by X-ray diffraction (XRD) technique, Atomic Force Microscopy (AFM) and Scanning electron microscopic (SEM) and antibacterial activity of TiO2 nanoparticles synthesized by L. crispatus were determined on the basis of minimum inhibitory concentration (MIC) values Effect of TiO2 nanoparticles on metallo-β lactamase and evaluation of combined effect between Antibiotics and synthesized TiO2 nanoparticles. Results: Fifty samples of urine were collected from patient suffering from recurrent UTI who had certain clinical symptoms. Forty four isolates (88%) were isolated and identified from fifty samples of urine, 13 isolates (29.5%) belonged to Escherichia coli which was the most predominant and 6 isolates (13.6%) belonged to Klebsiella pneumonia, 5 isolates (11.3%) belonged to Psudomonasaeruginosa , 2 isolates (4.5%) belonged to Acinetobacterbaumannii, one isolate belonged to M.morganii, 11 isolates (25%) belonged to S.aureusand 6 isolates (13.6%) belonged to S.saprophyticusMoreover , these isolates were tested for susceptibility to (13) antibiotics and for their ability to produce extended β –lactmase and metalo β- lactmaseenzymes. All isolates were resist to Ampicillin, Amoxicillin/ clavulanicacid and Aztreonam. Inhibitory effect of synthesized TiO2 nanoparticles was studied against growth and ESBL and Metallo beta lactamase of recurrent UTI causative bacteria. Results showed the MIC of TiO2 nanoparticles was (32) mg\ml against E. coli, M.morganii, A.baumanii and S.aureus, and (64) mg\ml against K.pneumoniae, P.aeruginosa and S.saprophyticus isolates.TiO2 nanoparticles showed inhibitory effect on metalloβ lactmase produced by E.coli, M.morganiiand A.baumanii., thereduction in inhibition zone diameter of MBL positive isolates of A.baumanii(1) and E.colifrom (2o to 25) mm after treatment with TiO2 nanoparticles compared to control without treatment with TiO2 nanoparticles (25-35,20- 30) mm respectively, while inhibition zone diameters of M.morganiiand A.baumanii (2) were(15,20),(10,20) mm respectively after treatment compared to control with inhibition zone diameters (23,30), (30,38) mm. The combined effect between Antibiotics and synthesized TiO2 nanoparticles was also investigated against multi resistant recurrent UTI causative bacteria. The antibacterial activities of some antibiotics such as Ampicillin and Gentamycin have been increased in the presence of TiO2 nanoparticle (subMIC) against all isolates. Conclusions: Titanium nanoparticles has more efficient antibacterial agent and can be useful in the treatment of infectious diseases caused bybacteria, the combination of antibiotics and metal nanoparticles could increase the antibiotics‘ efficacy against resistant pathogens. In addition, nanoparticle–antibiotic conjugates lower the amount of both agents in the dosage, which reduces harmfulness and increases antimicrobial properties.