Search

To date, no method has been developed to assess the distribution of mycotoxins on the surface of grains, or other plant material, and the depth of their penetration into the interior. The Infrared (IR) Laser Ablation-Remote-Electrospray Ionization (LARESI) platform coupled to a tandem mass spectrometer (MS/MS), measuring in selected reaction monitoring (SRM) mode, was employed for the targeted imaging of selected metabolites of Aspergillus fumigatus, including mycotoxins in biological objects for the first time. This methodology allowed for the localisation of grain metabolites and fungal metabolites of grain infected by this mould. The distribution of metabolites in spelt grain was differentiated: fumigaclavine C, fumitremorgin C, and fumiquinazoline D were located mainly in the embryo, brevianamide F in the seed coat, and fumagillin in the endosperm. The LARESI mass spectrometry imaging method can be used in the future for the metabolomic analysis of mould metabolites in various plants and agricultural products.

The aim of this study was to determine the cause of foxing on 19th century paper using omics methods, including metagenomics via high-throughput sequencing on the Illumina platform and metabolomics via high-resolution surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-ToF-MS) imaging using the gold nanoparticle-enhanced target (AuNPET) method. Metabolomic analysis as well as ninhydrin and amido black staining of paper samples showed the presence of proteinaceous substances in areas corresponding to foxing. Products of cellulose biodegradation and microbial metabolites, including yellow-coloured pigments (2-methyl-6-phytylquinol; delta-, gamma- and beta-tocopherols; 3-hydroxy-L-kynurenine) were detected. No difference in metal-ion content was seen between areas with and without foxing. Higher degrees of biodiversity were identified within foxing stains compared to foxing-free areas. Phoma sp. and Cladonia sp. moulds and Gluconobacter and Ralstonia bacteria were predominant in DNA samples obtained from foxing stains. The presence of these microbes has never previously been linked to the phenomenon of foxing. These studies indicate that foxing is a result of a combined effect of microbial action, paper-degradation by microbes, dye production, and chemical changes occurring in paper due to oxidation of cellulose metabolites.

Surface-assisted laser desorption/ionization mass spectrometry on gold nanoparticle enhanced target (AuNPET) technique was used for metabolomic analysis and secondary metabolites detection of two mould strains – Aspergillus versicolor and Penicillium chrysogenum in model conditions on microbiological malt extract agar medium. Results obtained with the use of AuNPET-based mass spectrometry technique were compared with traditional matrix-assisted laser desorption/ionization (MALDI) method based on α-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB) matrices. Gold nanoparticle enhanced target method enabled effective ionization of microbial cellular extract ingredients without interference from the matrix and also improved calibration of spectra resulting in the detection of much higher amount of characteristic metabolites for studied organisms than MALDI.

The aim of this study was to compare the metabolite profile of the moulds Alternaria, Aspergillus, Cladosporium, Penicillium, Stachybotrys, and Trichoderma under model conditions on MEA (malt extract agar) medium and carton-gypsum board (CGB). This was followed by comparison of the metabolite profile of each particular strain and of a mixed culture of all moulds together. Metabolome analysis was performed using a high-resolution surface assisted laser desorption/ionization time-of-flight mass spectrometry based on a gold nanoparticle-enhanced target (AuNPET SALDI-ToF-MS) imaging method. All water extracts tested showed the presence of compounds with molecular weights in the range of 80–2000 m/z (mass-to-charge ratio). On CGB, the results obtained showed the existence of 299–453 peaks with higher intensity and 351–487 peaks with lower intensity compared to the peaks obtained from MEA, depending on the mould species. A distinct change in the metabolite profile under the influence of the growth medium was observed for the mixed culture on the MEA medium. The profile showed ∼200 peaks with higher intensities of metabolites within m/z ranges 80–300 and 450–600; for the culture growing on CGB, the peak intensity was within m/z 300–450, and the number of peaks was 600–750. A new metabolomic methodology allowed the identification of 89 metabolites belonging to 48 metabolomic pathways. Compounds belonging to the citrate cycle and penicillin and cephalosporin biosynthesis pathways were identified on MEA. On CGB moulds, the metabolism changed to steroid, ubiquinone and other terpenoid-quinone biosynthesis.

Bioaerosol is a threat at workplaces, therefore the selection and safe use of filtering facepiece respirators (FFRs) is important in preventive activities. The aim of the study was to assess the survival of microorganisms on materials used for FFRs construction. The parameters for microorganism growth under model conditions were described using the Gompertz equation, model verification was also carried out using FFRs at the farmers&rsquo, workplaces. We found that the factors determining a high survival of microorganisms were as follows: moisture corresponding to the conditions of use and storage of FFRs at workplaces, the presence of sweat and organic dust, inorganic dust and addition of biocide in nonwovens limited the growth of microorganisms, resulting in a shortening of the stationary growth phase and decreased cell numbers (5&ndash, 6 log). Dust concentration at workplaces was higher than EU occupational exposure limit values and WHO recommendations for airborne particulate matter. Microbial contaminations of the air (103&ndash, 104 CFU/m3), settled dust (104&ndash, 106 CFU/g) and FFRs (105 CFU/4cm2) during the grain harvest were high, the main contamination being bacteria (actinomycetes, Pseudomonas fluorescens) and xerophilic fungi. A high correlation was found between the number of microorganisms and the weight of dust on FFRs (R2 = 0.93&ndash, 0.96).

Plant biomass processed in a heat and power plant is a source of airborne dust present in the working environment. The aim of this study was to assess the influence of various levels of organic dust content in the filter material on the viability of microorganisms in model conditions, taking into consideration the workplace environment and the physico-chemical as well as the microbiological characteristics of the dust particles. The possibility of a biofilm developing on the reusable filtering facepiece respirators (FFR) in standard use by heat and power plant workers for respiratory tract protection was also investigated. Standard reusable filtering half masks FFP3R and high-performance melt-blown fabrics were used in the study. The biomass dust characteristics were determined using an elementary analyser and a condensation particle counter with an electrostatic classifier. For quantitative analysis of microbiological pollution we used the plate count method with the microorganisms identified either using the API biochemical tests (for bacteria and yeasts) and the taxonomic key (for moulds). Biofilm development on the filtering half masks was assessed using SEM scanning microscopy. The viability of microorganisms (namely Escherichia coli ATCC 10536, Staphylococcus aureus ATCC 6538, Candida albicans ATCC 10231, Aspergillus niger ATCC 16404; Bacillus subtilis NCAIM 01644) on the filter materials with different dust content was measured using the quantitative method AATCC 100-2004 (2008). In the plant biomass processed at the heat and power plant, a high concentration of bacteria (2.30 × 10 7 CFU g −1 ) and a lower concentration of fungi (4.46 × 10 5 CFU g −1 ) were detected. It was determined to be a good environment for the growth of microorganisms (the carbon to nitrogen ratio was 48:1). The presence of dust and the development of microbiological biofilms on reusable FFR, which is used by the heat and power plant workers for processing the plant biomass in increased humidity conditions, were also confirmed. It was found that the viability of microorganisms on a filter material depends on the kind of microorganism and the dust content on the material. Model research showed that biomass dust in filter materials stimulates the growth of E. coli . Moreover, dust concentration above 21% inhibited the growth of S. aureus and C. albicans but had no significant influence on the growth of B. subtilis and A. niger .

The aim of this study was to characterise the microbial and metabolite diversity in samples of biodeteriorated 18th century silk from the burial crypts of the Church of St Francis of Assisi in Krakow (Poland). Highthroughput Illumina sequencing, surface-assisted laser desorption/ionisation mass spectrometry on a silver nanoparticle enhanced target (109Ag SALDI), and laser ablation-remote-electrospray ionisation-selected reaction monitoring-ambient mass spectrometry imaging (LARESI MSI) were employed. The use of high throughput Illumina sequencing helped obtain a broader picture of microbiocenosis compared to earlier studies. The 109Ag SALDI method enabled the qualitative analysis of the entire chemical compound profile of silk. A few hundred metabolites, including peptides, amino acids, urea and organic acids dominated by N-(3-oxododecanoyl)-L-homoserine, decanoylcholine, formiminoalanine and hexacosatrienoic acid were detected in archaeological silk, whereas about 100 metabolites, mainly represented by polymers, were detected in contemporary and restored silk. The novel ambient MS imaging method LARESI MSI, used for the first time for the archaeological silk biodeterioration analysis, allowed the direct detection and mapping of selected amino acids, historical dyes and dihydroxybenzoic acid (decomposition product of tannins) on the silk samples. Based on our results, the suitability of the tested methods should also be considered for other historical objects.

The aim of the study was to determine microbial and metabolites diversity on the samples of damaged mediaeval beeswax seals from the Archdiocese Archive of Gniezno, Poland. We employed high-throughput sequencing on the Illumina platform and 109Ag SALDI metabolome analysis for the first time to assess biodeterioration of the beeswax seals. Bacteria belonged to the genera Mesorhizobium, Delftia, Ralstonia, Burkholderia, Stenotrophomonas, Olsenella and fungi: Aspergillus, Penicillium, Aureobasidium, Chaetomium, Malassezia, and others microorganisms identified with lower relative abundance may be responsible for the biodeterioration of the seals analysed. Between 112 and 456 chemical compounds were identified on the seal samples. In all samples, fatty acids, mono- di- and triglycerides, hydrocarbons and lipids including glycerophosphocholines, glycerophosphoethanolamines, glycerophosphoserines, glycerophosphoglycerols, glycerophosphates, glycerophosphoinositols, ceramides and sphingomyelins, were detected. Chemical compounds including fatty oxyacids and metabolites of microbiological origin belonged to 20 metabolic pathways can be considered as indicators of biodeterioration of beeswax seals. The usefulness of new available methods - high-throughput sequencing on the Illumina platform and 109Ag SALDI in historical seals biodeterioration analysis was demonstrated.

Filtering nonwovens that constitute the base material for filtering facepiece respirators (FFRs) used for the protection of the respiratory system against bioaerosols may, in favourable conditions, promote the development of harmful microorganisms. There are no studies looking at the impact that different types of filtering nonwovens have on microorganism survival, which is an important issue for FFR producers and users. Five commercial filtering nonwovens manufactured using diverse textile technologies (i.e., needle-punching, melt-blown, spun-bonding) with different structural parameters and raw material compositions were used within our research. The survival of microorganisms on filtering nonwovens was determined for E. coli, S. aureus, B. subtilis bacteria, C. albicans yeast and A. niger mould. Samples of nonwovens were collected immediately after inoculum application (at 0 h) and after 4, 8, 24, 48, 72, and 96 h of incubation. The tests were carried out in accordance with the AATCC 100-1998 method. Survival depended strongly on microorganism species. E. coli and S. aureus bacteria grew the most on all nonwovens tested. The structural parameters of the nonwovens tested (mass per unit area and thickness) and contact angle did not significantly affect microorganism survival.

The aim of the study was to analyze the microbiological biodiversity of human foot skin with respect to factors such as age, gender, frequency of foot washing and physical activity, and to select indicator species to be considered when designing textile materials with antimicrobial properties used for sock and insole production. The experiment was carried out on a group of 40 people. The number of microorganisms was determined using culture-dependent methods. Biodiversity was determined using culture followed by genetic identification based on 16S rRNA gene sequencing (bacteria), ITS region (fungi), or using Illumina next-generation sequencing (in a group of eight selected individuals). The total bacterial number on women&rsquo, s feet was on average 1.0 &times, 106 CFU/cm2, and was not statistically significantly different than that of men&rsquo, s feet (mean 1.2 &times, 105 CFU/cm2). The number of bacteria, in most cases, decreased with age and with increased frequency of physical activity. The number of bacteria increased with diminishing feet-washing frequency, however, statistically significant differences were found between groups. The number of fungi was not significantly different amongst groups. Bacteria belonging to the phyla Firmicutes, Proteobacteria and Actinobacteria constituted the main microorganisms of the foot skin. Ascomycota and Basidiomycota predominated amongst the fungi. The presence of specific species varied in groups depending on the factors mentioned above. Two of the species identified were classified as pathogens (Neisseria flavescens and Aspergillus fumigatus). These findings suggest that it is necessary to extend the list of microorganisms tested on textiles with respect to hygienic properties.

The aim of the present study was to evaluate the relation between the chemical (analysis of elements and pH) and microbiological composition (culture and metagenomics analysis) of the dust at various workplaces (cement plant, composting plant, poultry farm, and cultivated area) and the cytotoxicity effect on the human adenocarcinoma lung epithelial adherent cell line A-549 (MTT assay test). Analysis of the Particulate Matter (PM) fraction showed that the dust concentration in cultivated areas exceeded the OELs. For the remaining workplaces examined, the dust concentration was lower than OELs limits. The number of microorganisms in the dust samples was 3.8 × 102–1.6 × 108 CFU/g bacteria and 1.5 × 102–6.5 × 106 CFU/g fungi. The highest number of microorganisms was noted for dust from cultivated areas (total number of bacteria, actinomycetes, P. fluorescens) and composting plants (xerophilic fungi and staphylococci), while the least number of microorganisms was observed for dust from cement plants. Many types of potentially pathogenic microorganisms have been identified, including bacteria, such as Bacillus, Actinomyces, Corynebacterium, Prevotella, Clostridium, and Rickettsia, and fungi, such as Alternaria, Cladosporium, Penicillium, and Aspergillus. The most cytotoxic to the human lung cell line A-549 was dust from cultivated areas (IC50 = 3.8 mg/mL after 72 h). The cytotoxicity of the tested dust samples depends on the PM concentration, the number of microorganisms, including potentially pathogenic genera, and the exposure time.