Aflatoxin production and in vitro toxicity of Aspergilli section Flavi isolated from air samples collected from different environments

Daniela Jakšić, S. Kocsubé, Ottó Bencsik, Anita Kecskeméti, A. Szekeres, Dubravko Jelić, Nevenka Kopjar, C. Vágvölgyi, J. Varga, Maja Šegvić Klarić

Research output: Article

Abstract

Aspergilli section Flavi, originally isolated from air samples collected from inhabited apartments (AP), unoccupied basements (BS), and processing facilities of a grain mill (GM), were analyzed for their potential to produce aflatoxin B 1 (AFB 1 ) on solid media. The isolates were further characterized with regard to their cytotoxic, genotoxic, and pro-inflammatory properties in vitro. Aspergilli were identified based on partial calmodulin (CaM) gene sequencing; the producing capacities of isolates were analyzed by HPLC/FLD and confirmed by genes in biosynthesis (aflR, norA, omtA). In the grain mill, the Aspergilli section Flavi (up to 1.3 × 10 6  cfu/m 3 ) dominated by AFB 1 -producing Aspergillus flavus (71%, 4.5–5254 ng/ml) which showed a serious health risk for workers. Living environments were not relevant sources of exposure. After 24 h, AFB 1 (1–100 μmol/l) reduced cell viability (MTT test) in both A549 cells and THP-1 macrophage-like cells without reaching IC 50 . In A549 cells, the extract of the AFB 1 -producing A. flavus significantly decreased cell viability but not below 50%. THP-1 macrophage-like cells were more sensitive to both extracts, but IC 50 was obtained only for the AFB 1 -producing strain (0.37 mg/ml; AFB 1 2.78 μmol/l). AFB 1 (1 and 10 μmol/l) induced significant DNA damage (tail intensity, alkaline comet assay) in A549 cells in contrast to Aspergilli extracts. AFB 1 elevated IL-6 and IL-8, while Aspergilli extracts increased IL-1β, TNF-α, and IL-17 release in THP-1 macrophages (ELISA). Chronic exposure to AFB 1 and/or other metabolites in airborne A. flavus from occupational environments may stimulate epithelial damage of airways accompanied by lowered macrophage viability.

Original languageEnglish
JournalMycotoxin Research
DOIs
Publication statusPublished - jan. 1 2019

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Aflatoxin B1
Aflatoxins
Aspergillus
Toxicity
Air
Macrophages
Aspergillus flavus
Cell Survival
Genes
Cells
In Vitro Techniques
Comet Assay
Interleukin-17
Health risks
Biosynthesis
Calmodulin
Metabolites
Cell Extracts
Interleukin-8
Interleukin-1

Keywords

    ASJC Scopus subject areas

    • Biotechnology
    • Microbiology
    • Toxicology

    Cite this

    Aflatoxin production and in vitro toxicity of Aspergilli section Flavi isolated from air samples collected from different environments. / Jakšić, Daniela; Kocsubé, S.; Bencsik, Ottó; Kecskeméti, Anita; Szekeres, A.; Jelić, Dubravko; Kopjar, Nevenka; Vágvölgyi, C.; Varga, J.; Šegvić Klarić, Maja.

    In: Mycotoxin Research, 01.01.2019.

    Research output: Article

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    abstract = "Aspergilli section Flavi, originally isolated from air samples collected from inhabited apartments (AP), unoccupied basements (BS), and processing facilities of a grain mill (GM), were analyzed for their potential to produce aflatoxin B 1 (AFB 1 ) on solid media. The isolates were further characterized with regard to their cytotoxic, genotoxic, and pro-inflammatory properties in vitro. Aspergilli were identified based on partial calmodulin (CaM) gene sequencing; the producing capacities of isolates were analyzed by HPLC/FLD and confirmed by genes in biosynthesis (aflR, norA, omtA). In the grain mill, the Aspergilli section Flavi (up to 1.3 × 10 6  cfu/m 3 ) dominated by AFB 1 -producing Aspergillus flavus (71{\%}, 4.5–5254 ng/ml) which showed a serious health risk for workers. Living environments were not relevant sources of exposure. After 24 h, AFB 1 (1–100 μmol/l) reduced cell viability (MTT test) in both A549 cells and THP-1 macrophage-like cells without reaching IC 50 . In A549 cells, the extract of the AFB 1 -producing A. flavus significantly decreased cell viability but not below 50{\%}. THP-1 macrophage-like cells were more sensitive to both extracts, but IC 50 was obtained only for the AFB 1 -producing strain (0.37 mg/ml; AFB 1 2.78 μmol/l). AFB 1 (1 and 10 μmol/l) induced significant DNA damage (tail intensity, alkaline comet assay) in A549 cells in contrast to Aspergilli extracts. AFB 1 elevated IL-6 and IL-8, while Aspergilli extracts increased IL-1β, TNF-α, and IL-17 release in THP-1 macrophages (ELISA). Chronic exposure to AFB 1 and/or other metabolites in airborne A. flavus from occupational environments may stimulate epithelial damage of airways accompanied by lowered macrophage viability.",
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    AU - Kecskeméti, Anita

    AU - Szekeres, A.

    AU - Jelić, Dubravko

    AU - Kopjar, Nevenka

    AU - Vágvölgyi, C.

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    AU - Šegvić Klarić, Maja

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