Viorica Railean-Plugaru

Study of silver nanoparticles synthesized by acidophilic strain of Actinobacteria isolated from the of Picea sitchensis forest soil

In the present work the acidophilic actinobacteria strain was used as a novel reducing agent for the cheap, green and single‐step synthesis of nanostructure silver particles. Structural, morphological and optical properties of the synthesized nanoparticles have been characterized by spectroscopy, dynamic light scattering and electron microscopy approach. The antimicrobial activity of silver nanoparticles against clinical strains such as Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis and Salmonella infantis alone and in combination with antibiotics were studied.

Antimicrobial properties of biosynthesized silver nanoparticles studied by flow cytometry and related techniques.

This work reports the effect of silver bionanoparticles (Bio(AgNPs) synthesized by Actinobacteria CGG 11n on selected Gram (+) and Gram (–) bacteria. Flow cytometry, classical antibiogram method and fluorescent microscopy approach was used for evaluation of antimicrobial activity of Bio(AgNPs) and their combination with antibiotics. Furthermore, the performed research specified the capacity of flow cytometry method as an alternative to the standard ones and as a complementary method to electromigration techniques. The study showed antibacterial activity of both BioAgNPs and the combination of antibiotics/BioAgNPs against all the tested bacteria strains in comparison with a diffusion, dilution and bioautographic methods. The synergistic effect of antibiotics/BioAgNPs combination (e.g. kanamycin, ampicillin, neomycin and streptomycin) was found to be more notable against Pseudomonas aeruginosa representing a prototype of multi‐drug resistant “superbugs” for which effective therapeutic options are very limited.

New approach for fast identification of cyclitols by MALDI-TOF mass spectrometry

INTRODUCTION Alfalfa (Medicago sativa L.) is the subject of many studies due to its numerous chemical constituents and beneficial properties. Among these constituents are cyclitols, which have attracted attention due to the variety of biological properties they have. OBJECTIVE A rapid and sensitive analytical procedure based on matrix-assisted laser desorption ionisation technique with time-of-flight and mass spectrometry (MALDI-TOF-MS) analysis was used for the first time for the identification of three cyclitols from different parts of alfalfa. METHODOLOGY Plant extracts were prepared and purified using Soxhlet extraction and solid-phase extraction (SPE). Then, samples were dissolved in α-cyano-4-hydroxycinnamic acid (HCCA) matrix, and subjected to MALDI-TOF-MS analysis. RESULTS The ion at m/z 524.0 was distributed in all standards and in leaves and stem extracts. In turn, the signal at m/z 335.1 was found in all standards and all alfalfa extracts. The ion at m/z144.1 was found just for d-chiro-inositol and distributed in all extracts. Both signals at m/z 265.9 and 250.0 were found only in l-chiro-inositol standard and the extract of stem. However, the ion at m/z 177.1 was found in d-pinitol standard and the extract of leaves. Based on molecular weights, information on fragment ions obtained by MALDI-TOF-MS, and the chemistry of cyclitols, we successfully identified three cyclitols (d-chiro-inositol, l-chiro-inositol, d-pinitol) in different parts of alfalfa (leaves, stem, flowers). CONCLUSION The obtained results in this study proved that MALDI-TOF-MS is a rapid, sensitive and very powerful tool for identification of cyclitols within plants and has the potential to differentiate between enantiomers.

Complex investigation of extraction techniques applied for cyclitols and sugars isolation from different species of Solidago genus

Cyclitols are phytochemicals naturally occurring in plant material, which attracted an increasing interest due to multiple medicinal attributes, among which the most important are the antidiabetic, antioxidant, and anticancer properties. Due to their valuable properties, sugars are used in the food industry as sweeteners, preservatives, texture modifiers, fermentation substrates, and flavoring and coloring agents. In this study, we report for the first time the quantitative analysis of sugars and cyclitols isolated from Solidago virgaurea L., which was used for the selection of the optimal solvent and extraction technique that can provide the best possible yield. Moreover, the quantities of sugars and cyclitols extracted from two other species, Solidago canadensis and Solidago gigantea, were investigated using the best extraction method and the most appropriate solvent. Comparative analysis of natural plant extracts obtained using five different techniques—maceration, Soxhlet extraction, pressurized liquid extraction, ultrasound‐assisted extraction, and supercritical fluid extraction—was performed in order to decide the most suitable, efficient, and economically convenient extraction method. Three different solvents were used. Analysis of samples has been performed by solid‐phase extraction for purification and pre‐concentration, followed by derivation and GC‐MS analysis. Highest efficiency for the total amount of obtained compounds has been reached by PLE, when water was used as a solvent. d‐pinitol amount was almost similar for every solvent and for all the extraction techniques involved.

Silver nanoparticles functionalized with ampicillin

The resistance of pathogenic bacteria to antibiotics has become a serious problem. The emphasis is placed on the development of new, effective antimicrobial strategies. One of them is the use of AgNPs in association with antibiotic drugs. The aim of this study was to obtain silver nanoparticles functionalized with ampicillin and to investigate the mechanism of binding antibiotics to nanoparticle using high‐performance liquid chromatography approach. To confirm the occurrence of silver nanoparticles functionalization, FTIR, MALDI‐TOF MS, and DLS analysis and zeta potential measurements were performed. Moreover we assessed the antibacterial activity of biologically synthesized nanoparticles functionalized with ampicillin against a range of gram (+) and gram (−) bacteria strains such as Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus epidermidis, and Escherichia coli.

Microbial Analysis of Escherichia coli ATCC, Lactobacteria and Saccharomyces cerevisiae Using Capillary Electrophoresis Approach.

Rapid detection and identification of microorganisms is a challenging and important aspect in many areas of our life, beginning with medicine, ending with industry. Unfortunately, classical methods of microorganisms identification are based on time-consuming and labor-intensive approaches. Screening techniques require rapid and cheap grouping of bacterial isolates; however, modern bioanalytics demands comprehensive bacterial studies on molecular level. The new approach to the rapid identification of bacteria is to use the electromigration techniques, especially capillary zone electrophoresis (CZE). CZE is an important technique used in the analysis of microorganisms. However, the analysis of microbial complexes using this technology still encounters several problems-uncontrolled aggregation and/or adhesion to the capillary surface. One way to resolve this issue is the CZE analysis of microbial cell with surface charge modification by bivalent metal ions (e.g., Ca(2+) aq, Zn aq). Under the above conditions, bacterial cells create compact aggregates, and fewer high-intensity signals are observed in electropherograms. The chapter presents the capillary electrophoresis of microbial aggregates approach with UV and one-dimensional intact cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (ICM MS) detection.

Microbiology neutralization of zearalenone using Lactococcus lactis and Bifidobacterium sp.

AbstractThe aim of the study was to neutralize zearalenone by lactic acid bacteria (LAB) such as Lactococcus lactis and Bifidobacterium sp. and investigate the mechanism of zearalenone (ZEA) binding. Neutralization of ZEA by LAB was confirmed by identification of binding kinetics and spectroscopic studies such as Fourier transform infrared spectroscopy (FT-IR) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The obtained results showed that the kinetic process of zearalenone binding to L. lactis is not homogeneous but is expressed with an initial rapid stage with about 90% of ZEA biosorption and with a much slower second step. In case of Bifidobacterium sp., the neutralization process is homogeneous; the main stage can be described with about 88% of ZEA biosorption. MALDI–TOF-MS measurements and FTIR analysis confirmed the uptake of zearalenone molecules by bacterial species. Moreover, the assessment of dead and live lactic acid bacteria cells after zearalenone treatment was performed using fluorescence microscopy. Graphical abstractMicrobiology neutralization of zearalenone using Lactococcus lactis and Bifidobacterium sp. was confirmed by identification of binding kinetics and spectroscopic studies such as FT-IR spectroscopy and MALDI-TOF-MS spectrometry. The mechanism of ZEA binding was also investigated.

Physicochemical study of natural fractionated biocolloid by asymmetric flow field-flow fractionation in tandem with various complementary techniques using biologically synthesized silver nanocomposites

AbstractAsymmetric flow field-flow fractionation coupled with use of ultraviolet–visible, multiangle light scattering (MALLS), and dynamic light scattering (DLS) detectors was used for separation and characterization of biologically synthesized silver composites in two liquid compositions. Moreover, to supplement the DLS/MALLS information, various complementary techniques such as transmission electron spectroscopy, Fourier transform infrared spectroscopy, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used. The hydrodynamic diameter and the radius of gyration of silver composites were slightly larger than the sizes obtained by transmission electron microscopy (TEM). Moreover, the TEM results revealed the presence of silver clusters and even several morphologies, including multitwinned. Additionally, MALDI-TOF MS examination showed that the particles have an uncommon cluster structure. It can be described as being composed of two or more silver clusters. The organic surface of the nanoparticles can modify their dispersion. We demonstrated that the variation of the silver surface coating directly influenced the migration rate of biologically synthesized silver composites. Moreover, this study proves that the fractionation mechanism of silver biocolloids relies not only on the particle size but also on the type and mass of the surface coatings. Because silver nanoparticles typically have size-dependent cytotoxicity, this behavior is particularly relevant for biomedical applications. Graphical abstractWorkflow for asymmetric flow field-flow fractionation of natural biologically synthesized silver nanocomposites