Ophiobolin A from Bipolaris oryzae Perturbs Motility and Membrane Integrities of Porcine Sperm and Induces Cell Death on Mammalian Somatic Cell Lines

Bencsik, Ottó; Papp, Tamás; Berta, M.; Zana, Annamária; Forgó, Péter; Dombi, György; Andersson, Maria A.; Salkinoja‐Salonen, Mirja; Vágvölgyi, Csaba; Szekeres, András

doi:10.3390/toxins6092857

Cited by 29 publications

(44 citation statements)

References 35 publications

(64 reference statements)

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“…The seeded board was incubated in a steel chamber (12·5 l) and sealed with a glass lid at RH 95% and 20 ± 1 °C. Biomass (2 mg) was scraped with a microscopic slide from the surface of the gypsum liner and dispersed in 40 μl ethanol, incubated at 60 °C for 10 min and then tested for toxicity to PK‐15 cells using inhibition of proliferation as endpoint and applying a fluorometric readout confirmed with microscopic examination as described in Bencsik et al (). Concentrations of conidia (2 μm) and hyphal fragments (larger than 0·1 μm) in biomass lysates and guttation droplets were calculated as the average from 10 microscopic fields by phase contrast microscope (Olympus CKX41 Tokyo Japan) with 400 × magnification.…”

Section: Methodsmentioning

confidence: 99%

Penicillium expansum strain isolated from indoor building material was able to grow on gypsum board and emitted guttation droplets containing chaetoglobosins and communesins A, B and D

et al. 2019

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AimsEmission of toxic metabolites in guttation droplets of common indoor fungi is not well documented. The aims of this study were (i) to compare mycotoxins in biomass and guttation droplets from indoor fungi from a building following health complaints among occupants, (ii) to identify the most toxic strain and to test if mycotoxins in guttation liquids migrated trough air and (iii) to test if toxigenic Penicillium expansum strains grew on gypsum board.Methods and ResultsBiomass suspensions and guttation droplets from individual fungal colonies representing Aspergillus, Chaetomium, Penicillium, Stachybotrys and Paecilomyces were screened toxic to mammalian cells. The most toxic strain, RcP61 (CBS 145620), was identified as Pen. expansum Link by sequence analysis of the ITS region and a calmodulin gene fragment, and confirmed by the Westerdijk Institute based on ITS and beta‐tubulin sequences. The strain was isolated from a cork liner, was able to grow on gypsum board and to produce toxic substances in biomass extracts and guttation droplets inhibiting proliferation of somatic cells (PK‐15, MNA, FL) in up to 20 000‐fold dilutions. Toxic compounds in biomass extracts and/or guttation droplets were determined by HPLC and LC‐MS. Strain RcP61 produced communesins A, B and D, and chaetoglobosins in guttation droplets (the liquid emitted from them) and biomass extracts. The toxins of the guttation droplets migrated c. 1 cm through air and condensed on a cool surface.ConclusionsThe mycotoxin‐containing guttation liquids emitted by Pen. expansum grown on laboratory medium exhibited airborne migration and were >100 times more toxic in bioassays than guttation droplets produced by indoor isolates of the genera Aspergillus, Chaetomium, Stachybotrys and Paecilomyces. Significance and Impact of the StudyToxic exudates produced by Pen. expansum containing communesins A, B and D, and chaetoglobosins were transferable by air. This may represent a novel mechanism of mycotoxin dispersal in indoor environment.

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Section: Methodsmentioning

confidence: 99%

Penicillium expansum strain isolated from indoor building material was able to grow on gypsum board and emitted guttation droplets containing chaetoglobosins and communesins A, B and D

et al. 2019

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“…Cytostatic toxicity was indicated if ≤5 mg mL −1 of the biomass inhibited the cell proliferation (ICP assay) of the PK-15 cells exposed for one to two days [13]. The toxicity assays performed with ethanol extract from pure fungal cultures were performed using porcine cells (sperms, somatic cell lines) as indicators according to [14][15][16]19,20].…”

Section: Rapid Screening Tests With Ex Vivo and In Vitro Assaysmentioning

confidence: 99%

Screening Mold Colonies by Using Two Toxicity Assays Revealed Indoor Strains of Aspergillus calidoustus Producing Ophiobolins G and K

Salo

Marik

Bencsik

et al. 2019

Toxins

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The occurrence and toxin production of the opportunistic pathogen Aspergillus calidoustus in Finnish buildings is not well documented in the literature. We tracked and identified four A. calidoustus colonies cultivated from indoor settled dusts and revealed the biological activities of crude biomass extracts. The toxic substances were identified as 6-epi-ophiobolin K, ophiobolin K, and ophiobolin G by high-performance liquid chromatography–mass spectrometry (HPLC-MS) based on chromatographic and mass spectrometry data (MS and MS/MS) on the crude extract of A. calidoustus strain MH34. A total of 29 fungal colonies collected from settled dust in an office room reported for indoor-air-related illnesses were screened for toxins that inhibited boar sperm motility in the BSMI (boar sperm motility inhibiting) assay and cell proliferation in the ICP (inhibition of cell proliferation) assays with PK-15 cells. Out of the 27 colonies tested as toxic, 12 colonies exhibiting conidiophores representative of the genera Chaetomium, Penicillium, and Paecilomyces were excluded from the study, while 13 colonies exhibited Aspergillus-like conidiophores. Biomass suspensions of these colonies were divided into two categories: Category 1 colonies (n = 4), toxic in the BSMI assay and the ICP assays, emitted blue fluorescence and grew at 37 °C; Category 2 colonies (n = 9), only toxic in the ICP assay, emitted orange fluorescence and exhibited limited or no growth at 37 °C. Colonies in Category 1 were pure-cultured, and the strains were named as MH4, MH21, MH34, MH36. Strain MH34 was identified as A. calidoustus by the internal transcribed spacer (ITS) sequences. Ethanol-soluble dry substances extracted from the biomass of the pure cultures exhibited a toxicological profile in the BSMI assay, SMID (sperm membrane integrity damage) assay, and ICP assay similar to that exhibited by pure ophiobolin A. Overall, the viable conidia of A. calidoustus in indoor settled dusts deserve attention when potentially hazardous mold species are monitored.

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“…Several studies applied these tests for screening bioactive secondary metabolites, e.g. Bipolaris oryzae, the causal agent of the brown spot disease in rice produces substances -identified later as ophiobolins -that can inhibit these cells in bioassays (Bencsik et al 2014).…”

Section: Introductionmentioning

confidence: 99%

Bioactive Peptaibols of Forest-Derived Trichoderma Isolates from Section Longibrachiatum

Marik

Szekeres

Andersson

et al. 2017

Soil Biological Communities and Ecosystem Resilience

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Filamentous fungi are producers of a large number of secondary metabo lites with wide spectra of biological effects. Among them, peptaibols represent a group of compounds produced mainly by members of the mycotrophic filamentous fungal genus Trichoderma. A simple peptaibol characterization strategy including purification and structural elucidation steps was applied to examine the peptaibol production of three strains from the Longibrachiatum section of genus Trichoderma, T. aethiopicum TUCIM 1817, T. novae-zelandiae TUCIM 4158 and T. pseudokon ingii TUCIM 1277, all deriving from natural forest habitats (disturbed semiforest, native Notophagus forest and the bark of Beilschmiedia tawa, respectively). After the solid phase clean-up of culture extracts, mass spectrometric analysis of peptai bols produced by the examined strains was performed by on-line reversed-phase

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Ophiobolin A from Bipolaris oryzae Perturbs Motility and Membrane Integrities of Porcine Sperm and Induces Cell Death on Mammalian Somatic Cell Lines

Cited by 29 publications

References 35 publications

Penicillium expansum strain isolated from indoor building material was able to grow on gypsum board and emitted guttation droplets containing chaetoglobosins and communesins A, B and D

Penicillium expansum strain isolated from indoor building material was able to grow on gypsum board and emitted guttation droplets containing chaetoglobosins and communesins A, B and D

Screening Mold Colonies by Using Two Toxicity Assays Revealed Indoor Strains of Aspergillus calidoustus Producing Ophiobolins G and K

Bioactive Peptaibols of Forest-Derived Trichoderma Isolates from Section Longibrachiatum

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