Contact Mode Atomic Force Microscopy as a Rapid Technique for Morphological Observation and Bacterial Cell Damage Analysis.

Electron microscopy is one of the tools required to characterize cellular structures. However, the procedure is complicated and expensive due to the sample preparation for observation. Atomic force microscopy (AFM) is a very useful characterization technique due to its high resolution in three dimensions and because of the absence of any requirement for vacuum and sample conductivity. AFM can image a wide variety of samples with different topographies and different types of materials. AFM provides high-resolution 3D topography information from the angstrom level to the micron scale. Unlike traditional microscopy, AFM uses a probe to generate an image of the surface topography of a sample. In this protocol, the use of this type of microscopy is suggested for the morphological and cell damage characterization of bacteria fixed on a support. Strains of Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), and Pseudomonas hunanensis (isolated from garlic bulb samples) were used. In this work, bacterial cells were grown in specific culture media. To observe cell damage, Staphylococcus aureus and Escherichia coli were incubated with different concentrations of nanoparticles (NPs). A drop of bacterial suspension was fixed on a glass support, and images were taken with AFM at different scales. The images obtained showed the morphological characteristics of the bacteria. Further, employing AFM, it was possible to observe the damage to the cellular structure caused by the effect of NPs. Based on the images obtained, contact AFM can be used to characterize the morphology of bacterial cells fixed on a support. AFM is also a suitable tool for the investigation of the effects of NPs on bacteria. Compared to electron microscopy, AFM is an inexpensive and easy-to-use technique.

Amino Acid Complexes of Zirconium in a Carbon Composite for the Efficient Removal of Fluoride Ions from Water.

Amino acid complexes of zirconia represent an entirely new class of materials that were synthesized and studied for the first time for the decontamination of fluoride ion containing aqueous solutions. Glutamic and aspartic acid complexes of zirconia assembled with thin carbon (stacked graphene oxide) platelets deriving from graphite oxide (GO) were synthesized by a two-step method to prepare adsorbents. The characterization of the complexes was carried out using infrared spectroscopy to determine the functional groups and the types of interaction between the composites and fluoride ions. To reveal the mechanisms and extent of adsorption, two types of batch adsorption measurements were performed: (i) varying equilibrium fluoride ion concentrations to construct adsorption isotherms at pH = 7 in the absence of added electrolytes and (ii) using fixed initial fluoride ion concentrations (10 mg/L) with a variation of either the pH or the concentration of a series of salts that potentially interfere with adsorption. The experimental adsorption isotherms were fitted by three different theoretical isotherm equations, and they are described most appropriately by the two-site Langmuir model for both adsorbents. The adsorption capacities of Zr-glutamic acid-graphite oxide and Zr-aspartic acid-graphite oxide are 105.3 and 101.0 mg/g, respectively. We found that two distinct binding modes are combined in the Zr-amino acid complexes: at low solution concentrations, F- ions are preferentially adsorbed by coordinating to the surface Zr species up to a capacity of ca. 10 mg/g. At higher concentrations, however, large amounts of fluoride ions may undergo anion exchange processes and physisorption may occur on the positively charged ammonium moieties of the interfacially bound amino acid molecules. The high adsorption capacity and affinity of the studied dicarboxylate-type amino acids demonstrate that amino acid complexes of zirconia are highly variable materials for the safe and efficient capture of strong Lewis base-type ions such as fluoride.

Two-Step Triethylamine-Based Synthesis of MgO Nanoparticles and Their Antibacterial Effect against Pathogenic Bacteria.

Magnesium oxide nanoparticles (MgO NPs) were obtained by the calcination of precursor microparticles (PM) synthesized by a novel triethylamine-based precipitation method. Scanning electron microscopy (SEM) revealed a mean size of 120 nm for the MgO NPs. The results of the characterizations for MgO NPs support the suggestion that our material has the capacity to attack, and have an antibacterial effect against, Gram-negative and Gram-positive bacteria strains. The ability of the MgO NPs to produce reactive oxygen species (ROS), such as superoxide anion radicals (O2•-) or hydrogen peroxide (H2O2), was demonstrated by the corresponding quantitative assays. The MgO antibacterial activity was evaluated against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria, with minimum inhibitory concentrations (MICs) of 250 and 500 ppm on the microdilution assays, respectively. Structural changes in the bacteria, such as membrane collapse; surface changes, such as vesicular formation; and changes in the longitudinal and horizontal sizes, as well as the circumference, were observed using atomic force microscopy (AFM). The lipidic peroxidation of the bacterial membranes was quantified, and finally, a bactericidal mechanism for the MgO NPs was also proposed.