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solution was heated for 8 min until it turned a pale yellow color. The solution was cooled to room temperature and centrifuged at 3,290 × g for 20 min. Then, 90% of the supernatant was removed from the centrifuged solution. The final concentration of Ag NP solution was 9.2 mM. The size of ZnO NPs and Ag NPs were determined by analyzing transmission electron microscopy (TEM) images using ImageJ software available at http://rsb.info.nih.gov/ij/. 3.3 Effect of ZnO NPs on the growth of E. coli K-12, L. acidophilus ADH, and B. animalis Bif-6 All three strains, incubated overnight in respective broth media, TSBY and MRS 0, were inoculated into broth media containing different concentrations (0, 12, 16, 20 mM) of ZnO NP suspensions, and 1% of NP-free solution. The NP-free solution was prepared by filtering ZnO NP suspensions through an anodisc inorganic membrane with a 20 nm pore size (Whatman Inc., Clifton, NJ, USA). Both ZnO NP suspensions and the NP-free solution were added to the broth media before autoclaving. After inoculating the cultures to the broth media, TSB or MRS, containing ZnO NP suspensions and NP-free solution, were incubated at 37 oC in a shaking incubator. The tubes inoculated with L. acidophilus ADH and B. animalis Bif-6 were placed in an anaerobic jar with GasPak (BD, GasPak EZ Anaerobe Container System) and incubated in a shaking incubator (Lab-Line 3528 Shaking incubator). The reason for using a shaking incubator was to avoid the aggregation of ZnO NPs in the broth and to allow a consistent contact between the bacterial cells and ZnO NPs. The samples were diluted 19PDF Image | ZINC OXIDE AND SILVER NANOPARTICLES ON INTESTINAL BACTERIA
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