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      Antibacterial activity of silver and zinc nanoparticles against Vibrio cholerae and enterotoxic Escherichia coli

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          Abstract

          Vibrio cholerae and enterotoxic Escherichia coli (ETEC) remain two dominant bacterial causes of severe secretory diarrhea and still a significant cause of death, especially in developing countries. In order to investigate new effective and inexpensive therapeutic approaches, we analyzed nanoparticles synthesized by a green approach using corresponding salt (silver or zinc nitrate) with aqueous extract of Caltropis procera fruit or leaves. We characterized the quantity and quality of nanoparticles by UV–visible wavelength scans and nanoparticle tracking analysis. Nanoparticles could be synthesized in reproducible yields of approximately 10 8 particles/ml with mode particles sizes of approx. 90–100 nm. Antibacterial activity against two pathogens was assessed by minimal inhibitory concentration assays and survival curves. Both pathogens exhibited similar resistance profiles with minimal inhibitory concentrations ranging between 5 × 10 5 and 10 7 particles/ml. Interestingly, zinc nanoparticles showed a slightly higher efficacy, but sublethal concentrations caused adverse effects and resulted in increased biofilm formation of V. cholerae. Using the expression levels of the outer membrane porin OmpT as an indicator for cAMP levels, our results suggest that zinc nanoparticles inhibit adenylyl cyclase activity. This consequently deceases the levels of this second messenger, which is a known inhibitor of biofilm formation. Finally, we demonstrated that a single oral administration of silver nanoparticles to infant mice colonized with V. cholerae or ETEC significantly reduces the colonization rates of the pathogens by 75- or 100-fold, respectively.

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          Most cited references108

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          A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria.

          J N Eloff (1998)
          Agar diffusion techniques are used widely to assay plant extracts for antimicrobial activity, but there are problems associated with this technique. A micro-dilution technique was developed using 96-well microplates and tetrazolium salts to indicate bacterial growth. p-Iodonitrotetrazolium violet [0.2 mg/ml] gave better results than tetrazolium red or thiazolyl blue. The method is quick, worked well with Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli and with non-aqueous extracts from many different plants. The method gave reproducible results; required only 10-25 microliters of extract to determine minimal inhibitory concentrations, distinguished between microcidal and microstatic effects, and provided a permanent record of the results. Using S. aureus, and a Combretum molle extract, the technique was 32 times more sensitive than agar diffusion techniques and was not sensitive to culture age of the test organism up to 24 hours. The S. aureus culture could be stored up to 10 days in a cold room with little effect on the assay results. This method was useful in screening plants for antimicrobial activity and for the bioassay-guided isolation of antimicrobial compounds from plants. MIC values determined for sulfisoxazole, norfloxacin, gentamicin, and nitrofuratoin were similar to values indicated in the literature but values obtained with trimethroprim and ampicillin were higher with some bacteria.
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            Toxicological impact studies based on Escherichia coli bacteria in ultrafine ZnO nanoparticles colloidal medium.

            We report here preliminary studies of biocidal effects and cellular internalization of ZnO nanoparticles on Escherichia coli bacteria. ZnO nanoparticles were synthesized in di(ethylene glycol) (DEG) medium by forced hydrolysis of ionic Zn2+ salts. Particle size and shape were controlled by addition of small molecules and macromolecules such as tri-n-octylphosphine oxide, sodium dodecyl sulfate, polyoxyethylene stearyl ether, and bovine serum albumin. Transmission electron microscopy (TEM) and X-ray diffraction analyses were used to characterize particle structure, size, and morphology. Bactericidal tests were performed in Luria-Bertani medium on solid agar plates and in liquid systems with different concentrations of small and macromolecules and also with ZnO nanoparticles. TEM analyses of bacteria thin sections were used to study biocidal action of ZnO materials. The results confirmed that E. coli cells after contact with DEG and ZnO were damaged showing a Gram-negative triple membrane disorganization. This behavior causes the increase of membrane permeability leading to accumulation of ZnO nanoparticles in the bacterial membrane and also cellular internalization of these nanoparticles.
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              Rapid biological synthesis of silver nanoparticles using plant leaf extracts.

              Five plant leaf extracts (Pine, Persimmon, Ginkgo, Magnolia and Platanus) were used and compared for their extracellular synthesis of metallic silver nanoparticles. Stable silver nanoparticles were formed by treating aqueous solution of AgNO(3) with the plant leaf extracts as reducing agent of Ag(+) to Ag(0). UV-visible spectroscopy was used to monitor the quantitative formation of silver nanoparticles. Magnolia leaf broth was the best reducing agent in terms of synthesis rate and conversion to silver nanoparticles. Only 11 min was required for more than 90% conversion at the reaction temperature of 95 degrees C using Magnolia leaf broth. The synthesized silver nanoparticles were characterized with inductively coupled plasma spectrometry (ICP), energy dispersive X-ray spectroscopy (EDS), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and particle analyzer. The average particle size ranged from 15 to 500 nm. The particle size could be controlled by changing the reaction temperature, leaf broth concentration and AgNO(3) concentration. This environmentally friendly method of biological silver nanoparticles production provides rates of synthesis faster or comparable to those of chemical methods and can potentially be used in various human contacting areas such as cosmetics, foods and medical applications.
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                Author and article information

                Contributors
                Journal
                Int J Med Microbiol
                Int. J. Med. Microbiol
                International Journal of Medical Microbiology
                Urban & Fischer Verlag
                1438-4221
                1618-0607
                1 January 2015
                January 2015
                : 305
                : 1
                : 85-95
                Affiliations
                [a ]University of Graz, Institute of Molecular Biosciences, BioTechMed-Graz, Humboldtstrasse 50, A-8010 Graz, Austria
                [b ]South Valley University, Faculty of Science, Qena, Egypt
                [c ]Institute of Biophysics, Medical University of Graz, BioTechMed-Graz, Schmiedlstraße 6, 8042 Graz, Austria
                [d ]Institute for Chemistry, Analytical Chemistry, University of Graz, BioTechMed-Graz, 8010 Graz, Austria
                Author notes
                [* ]Corresponding author. Tel.: +43 0316 380 1970; fax: +43 0316 380 9019. stefan.schild@ 123456uni-graz.at
                [1]

                These authors contributed equally to this work.

                Article
                S1438-4221(14)00153-2
                10.1016/j.ijmm.2014.11.005
                4300426
                25466205
                b139b463-a1e6-46aa-a547-9cd50a546749
                © 2014 The Authors

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).

                History
                : 5 June 2014
                : 30 October 2014
                : 4 November 2014
                Categories
                Article

                Microbiology & Virology
                caltropis procera,nanoparticles,in vivo,colonization,infant mouse model,biofilm,minimal inhibitory concentration,survival curve,therapeutic agent,antimicrobial activity

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