Determination of ergosterol in organic dust by gas chromatography-mass spectrometry

Axelsson, Bengt; Saraf, Anita; Larsson, Lennart

doi:10.1016/0378-4347(94)00553-h

Cited by 85 publications

(66 citation statements)

References 18 publications

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“…The presented ERG analysis with the application of HPLC was used to determine fungal biomass especially in, e.g., naturally contaminated cereal grain and in strongly contaminated cereal grain (Seitz et al 1977;Miedaner & Perkowski 1996;Šrobárová & Pavlová 2002;Wiśniewska & Buśko 2005), soil (Ruzicka et al 2000), feeds (Müller & Schwadorf 1993), construction materials (Miller et al 1988), dusts and aerosols (Axelsson et al 1995;Saraf et al 1997;Lau et al 2006), as well as foodstuffs (De Sio et al 2000;Kadakal et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Concentration of ergosterol in small-grained naturally contaminated and inoculated cereals

Perkowski¹,

Buśko²,

Stuper³

et al. 2008

Biologia

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Ergosterol (ERG) is a major sterol constituent of most fungi. Its concentration is negligible in higher plants, but can be used as a chemical marker of the presence of fungal contaminations. In this study, ERG concentration was assessed in randomly collected samples of naturally contaminated grain (wheat, barley and oat) and in samples of grain (wheat, barley, triticale and oat) harvested after inoculation of heads with conidia of different Fusarium species. Wheat samples were analysed at three stages of grain development. The lowest ERG concentration was found in non-inoculated samples at the first stage of grain development. This concentration was increasing with grain ripening. In naturally contaminated samples collected after harvest, ERG concentration was lower in wheat than in barley and oat. ERG concentrations in inoculated samples varied significantly, but were always significantly higher than in naturally contaminated samples. In the above cereal samples it was much lower than the levels assayed in laboratory cultures inoculated with fungi from genus Fusarium. The content of ERG was also analyzed in milling products of small-grained cereals and other foodstuffs, where a considerable variation was observed. The lowest ERG amounts were assayed in flours with a high degree of purification, while the highest ones in case of flours and products with a low purification rate. The results indicate the potential application of HPLC combined with microwave-assisted extraction both when assaying samples with low ERG concentrations (naturally contaminated) and those characterized with high contents of fungal biomass (strongly infected, artificially inoculated). It also facilitates analyses of fungal biomass in technological processes, where results may be expected to vary considerably.

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Section: Introductionmentioning

confidence: 99%

Concentration of ergosterol in small-grained naturally contaminated and inoculated cereals

Perkowski¹,

Buśko²,

Stuper³

et al. 2008

Biologia

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“…Muramic acid is a unique marker of peptidoglycan (2,8,9,15), which is the cross-linked macromolecular structure responsible for the rigidity of bacterial cell walls and is sometimes referred to as "gram-positive bacterial endotoxin" (27). Ergosterol is a common fungal membrane lipid that is widely used as a marker of fungal biomass, although the concentration depends on the species and growth conditions (1,6).…”

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confidence: 99%

Characterization of the Microbial Community in Indoor Environments: a Chemical-Analytical Approach

Sebastian

Larsson

2003

Appl Environ Microbiol

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An integrated procedure is presented whereby gas chromatography-ion trap mass spectrometry is used to determine chemical markers of gram-negative bacterial lipopolysaccharide (3-hydroxy fatty acids with 10 to 18 carbon atoms), gram-positive bacteria (branched-chain fatty acids with 15 and 17 carbon atoms), bacterial peptidoglycan (muramic acid), and fungal biomass (ergosterol) in samples of settled house dust. A hydrolysate of 13 C-labeled cyanobacterial cells is used as an internal standard for the first three markers. These analyses require two dust samples, one for 3-OH fatty acids, branched-chain fatty acids, and muramic acid and another for ergosterol. The method may be used to characterize microbial communities in environmental samples.Inhalation of airborne microorganisms in indoor environments has been associated with the development of respiratory disorders. Substances such as endotoxins (lipopolysaccharides [LPS]) (14,17,18,28), peptidoglycan (9, 12), and various fungal components and products (3,6,21) are among the suspected causative agents. Culture-based methods are suitable for detection of culturable infectious agents and allow species identification; however, it is widely agreed that only a small fraction (0.1 to 10%) of the total microbial flora in an indoor environment is currently culturable (29). Direct microscopy is at best semiquantitative. Although the Limulus amebocyte lysate test is extremely sensitive to endotoxin and can detect glucans, it measures bioactivity rather than absolute amounts and its reproducibility and specificity have been questioned (5). Nucleic acid-based methods including PCR are very specific, and by using broad-range (universal) probes and primers sets based on 16S rDNA (20) and 18S rRNA (31), most bacteria and fungi present in a sample can be identified. To date, such universal probes and primers have not been used to characterize the microbiology of indoor environments; rather, PCR has been used to detect specific fungi and bacteria in such environments (4,11,22).In our laboratory we have developed and applied gas chromatography-mass spectrometry (GC-MS) methods to determine biomarker molecules in complex matrices including organic dust. Microorganisms contain unique compounds not found elsewhere in nature, which can be used as chemical markers of larger, bioactive structures (7). Endotoxins (LPS) are major constituents of the outer membrane of gram-negative bacteria. A backbone of lipid A, the toxic component of the LPS molecule, carries in general 4 mol of unique 3-hydroxy fatty acids (3-OH FAs) (23-26). Certain branched-chain FAs are found in most gram-positive bacteria (13,30). Muramic acid is a unique marker of peptidoglycan (2,8,9,15), which is the cross-linked macromolecular structure responsible for the rigidity of bacterial cell walls and is sometimes referred to as "gram-positive bacterial endotoxin" (27). Ergosterol is a common fungal membrane lipid that is widely used as a marker of fungal biomass, although the concentration depends on the species and ...

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“…The lowest detectable cytokine concentrations were about 5 pg/ml. 3 /liter, and 0.5 g of PO 4 /liter. The contents of these nutrients were still considerably lower in the DSG.…”

Section: Discussionmentioning

confidence: 99%

“…The determination of the ergosterol content of S. chartarum grown on different plasterboards was based on previously published methods (4,28). Briefly, ergosterol standards (Sigma, St. Louis, Mo.)…”

Section: Methodsmentioning

confidence: 99%

Effect of Plasterboard Composition on Stachybotrys chartarum Growth and Biological Activity of Spores

Murtoniemi¹,

Nevalainen²,

Hirvonen³

2003

Appl Environ Microbiol

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The effects of plasterboard composition on the growth and sporulation of Stachybotrys chartarum as well as on the inflammatory potential of the spores were studied. S. chartarum was grown on 13 modified plasterboards under saturated humidity conditions. The biomass was estimated by measuring the ergosterol content of the S. chartarum culture while the spore-induced cytotoxicity and production of nitric oxide (NO), tumor necrosis factor alpha (TNF-␣), and interleukin-6 in mouse macrophages was used to illustrate the bioactivity of spores. The ergosterol content of S. chartarum correlated with the number of spores collected from plasterboards. The growth and sporulation decreased compared to that of the reference board in those cases where (i) the liner was treated with biocide, (ii) starch was removed from the plasterboard, or (iii) desulfurization gypsum was used in the core. Spores collected from all the plasterboards were toxic to the macrophages. The biocide added to the core did not reduce the growth; in fact, the spores collected from that board evoked the highest cytotoxicity. The conventional additives used in the core had inhibitory effects on growth. Recycled plasterboards used in the core and the board lacking the starch triggered spore-induced TNF-␣ production in macrophages. In summary, this study shows that the growth of a strain of S. chartarum on plasterboard and the subsequent bioactivity of spores were affected by minor changes to the composition of the core or liners, but it could not be totally prevented without resorting to the use of biocides. However, incomplete prevention of microbial growth by biocides even increased the cytotoxic potential of the spores.

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Determination of ergosterol in organic dust by gas chromatography-mass spectrometry

Cited by 85 publications

References 18 publications

Concentration of ergosterol in small-grained naturally contaminated and inoculated cereals

Concentration of ergosterol in small-grained naturally contaminated and inoculated cereals

Characterization of the Microbial Community in Indoor Environments: a Chemical-Analytical Approach

Effect of Plasterboard Composition on Stachybotrys chartarum Growth and Biological Activity of Spores

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