Analysis of bacteria associated with honeys of different geographical and botanical origin using two different identification approaches: MALDI-TOF MS and 16S rDNA PCR technique

Pomastowski, Paweł; Złoch, Michał; Rodzik, A.; Ligor, Magdalena; Kostrzewa, Markus; Buszewski, Bogusław

doi:10.1371/journal.pone.0217078

Cited by 55 publications

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References 59 publications

(70 reference statements)

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“…The probiotic strain was isolated from sweet whey at pH > 6 (Dairy Cooperative in Drzycim, Poland) according to the previous protocol [ 26 ]. The molecular identification of bacterial strain was performed by MALDI-TOF-MS identification by the ultrafleXtreme mass spectrometer (Bruker Daltonics, Hamburg, Germany) using formic acid-acetonitrile extraction and BioTyper identification [ 26 ]. Finally, the isolated strains of L. paracasei LPC20 were deposited in the Polish Collection of Microorganisms (PCM) under deposit no.…”

Section: Methodsmentioning

confidence: 99%

Zinc Oxide Nanocomposites—Extracellular Synthesis, Physicochemical Characterization and Antibacterial Potential

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This research presents, for the first time, the potential of the Lactobacillus paracasei LC20 isolated from sweet whey as a novel, effective and accessible source for post-cultured ZnO nanocomposites synthesis. The obtained nanocomposites were subjected to comprehensive characterization by a broad spectrum of instrumental techniques. Results of spectroscopic and microscopic analysis confirmed the hexagonal crystalline structure of ZnO in the nanometer size. The dispersion stability of the obtained nanocomposites was determined based on the zeta potential (ZP) measurements—the average ZP value was found to be −29.15 ± 1.05 mV in the 7–9 pH range. The ZnO nanocomposites (NCs) demonstrated thermal stability up to 130 °C based on the results of thermogravimetric TGA/DTG) analysis. The organic deposit on the nanoparticle surface was recorded by spectroscopic analysis in the infrared range (FT-IR). Results of the spectrometric study exhibited nanostructure-assisted laser desorption/ionization effects and also pointed out the presence of organic deposits and, what is more, allowed us to identify the specific amino acids and peptides present on the ZnO NCs surfaces. In this context, mass spectrometry (MS) data confirmed the nano-ZnO formation mechanism. Moreover, fluorescence data showed an increase in fluorescence signal in the presence of nanocomposites designed for potential use as, e.g., biosensors. Despite ZnO NCs’ luminescent properties, they can also act as promising antiseptic agents against clinically relevant pathogens. Therefore, a pilot study on the antibacterial activity of biologically synthesized ZnO NCs was carried out against four strains (Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa) by using MIC (minimal inhibitory concentration). Additionally, the colony forming units (CFU) assay was performed and quantified for all bacterial cells as the percentage of viable cells in comparison to a control sample (untreated culture) The nanocomposites were effective among three pathogens with MIC values in the range of 86.25–172.5 μg/mL and showed potential as a new type of, e.g., medical path or ointment formulation.

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Section: Methodsmentioning

confidence: 99%

Zinc Oxide Nanocomposites—Extracellular Synthesis, Physicochemical Characterization and Antibacterial Potential

et al. 2020

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“…After air-drying, the sample spot was overlaid with 1 μL of respective matrix solution: (1) α-Cyano-4-hydroxycinnamic acid (HCCA): 10 mg/mL in standard solvent solution (50% ACN, 47.5% water and 2.5% trifluoro acetic acid); (2) 2,5-dihydroxybenzoic acid (DHB): 50 mg/mL or (3) SuperDHB (9:1 ( w / w ) mixture of DHB and 2-hydroxy-5-methoxybenzoic acid, respectively): 50 mg/mL in standard solvent solution. MALDI Target plates with the samples were analyzed using ultrafleXtreme MALDI–TOF/TOF mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany) equipped with the smartbeam-II laser–positive mode according to the procedure described in detail in a previous work [ 56 ]. Spectra were collected manually using manufacturer software, flexControl, and the following parameters: m / z range: 2000–20,000, acceleration voltage at 25 kV, global attenuator offset at 20% and attenuator offset at 34% and its range at 34%, laser power at 40%, 500 shots in-one-single spectra to frequency 2500.…”

Section: Methodsmentioning

confidence: 99%

Study on Molecular Profiles of Staphylococcus aureus Strains: Spectrometric Approach

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Staphylococcus aureus remains a major health problem responsible for many epidemic outbreaks. Therefore, the development of efficient and rapid methods for studying molecular profiles of S. aureus strains for its further typing is in high demand. Among many techniques, matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI TOF MS) represents a timely, cost-effective, and reliable strain typing approach, which is still rarely used due to insufficient knowledge about the impact of sample preparation and analysis conditions on the molecular profiles and strain classification efficiency of S. aureus. The aim of this study was to evaluate the effect of the culture conditions and matrix type on the differentiation of molecular profiles of various S. aureus strains via the MALDI TOF MS analysis and different computational methods. The analysis revealed that by changing the culture conditions, matrix type, as well as a statistical method, the differentiation of S. aureus strains can be significantly improved. Therefore, to accelerate the incorporation of the MALDI-based strain typing in routine laboratories, further studies on the standardization and searching of optimal conditions on a larger number of isolates and bacterial species are of great need.

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“…A similar conclusion can be made for C 4 (11)3 and C 4 (11)2 isolates phylogenetically related to B. coreaensis and C. ramotenellum respectively, whose production and secretion of xylanases and laccases is one of their distinctive metabolic traits 37 , 40 , 44 . Further, Bacillus [C 4 (13)2], Staphylococci [C 5 (13)2 and C 4 (15)1)], and Micrococci [C 2 (11)1 and C 5 (13)1] species are among the most persistent strains capable of metabolizing honey and beeswax 51 , while M. chokoriensis [C 7 (11)1] and C. ramotenellum [C 4 (11)2] are proficient in degrading gelatine and starch, which were two of the most used binders in Japanese tradition 15 , 27 .…”

Section: Resultsmentioning

confidence: 99%

A combined physical–chemical and microbiological approach to unveil the fabrication, provenance, and state of conservation of the Kinkarakawa-gami art

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et al. 2020

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Kinkarakawa-gami wallpapers are unique works of art produced in Japan between 1870 and 1905 and exported in European countries, although only few examples are nowadays present in Europe. So far, neither the wallpapers nor the composing materials have been characterised, limiting the effective conservation–restoration of these artefacts accounting also for the potential deteriogen effects of microorganisms populating them. In the present study, four Kinkarakawa-gami wallpapers were analysed combining physical–chemical and microbiological approaches to obtain information regarding the artefacts’ manufacture, composition, dating, and their microbial community. The validity of these methodologies was verified through a fine in blind statistical analysis, which allowed to identify trends and similarities within these important artefacts. The evidence gathered indicated that these wallpapers were generated between 1885 and 1889, during the so-called industrial production period. A wide range of organic (proteinaceous binders, natural waxes, pigments, and vegetable lacquers) and inorganic (tin foil and pigments) substances were used for the artefacts’ manufacture, contributing to their overall complexity, which also reflects on the identification of a heterogeneous microbiota, often found in Eastern environmental matrices. Nevertheless, whether microorganisms inhabiting these wallpapers determined a detrimental or protective effect is not fully elucidated yet, thus constituting an aspect worth to be explored to deepen the knowledge needed for the conservation of Kinkarakawa-gami over time.

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Analysis of bacteria associated with honeys of different geographical and botanical origin using two different identification approaches: MALDI-TOF MS and 16S rDNA PCR technique

Cited by 55 publications

References 59 publications

Zinc Oxide Nanocomposites—Extracellular Synthesis, Physicochemical Characterization and Antibacterial Potential

Zinc Oxide Nanocomposites—Extracellular Synthesis, Physicochemical Characterization and Antibacterial Potential

Study on Molecular Profiles of Staphylococcus aureus Strains: Spectrometric Approach

A combined physical–chemical and microbiological approach to unveil the fabrication, provenance, and state of conservation of the Kinkarakawa-gami art

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