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Fluorescent Viability Stains to Probe the Metabolic Status of Aflatoxigenic Fungus in Dual Culture of Aspergillus flavus and Pichia anomala

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Abstract

The metabolic activity of the aflatoxigenic fungus, Aspergillus flavus co-cultured with the biocontrol yeast, Pichia anomala was examined using several viability stains. Both the FUN-1 stain and the combined use of DiBAC4(5) with CDFA-AM stains were applied in this study. The results suggest that the ATP-generating system in A. flavus was inactivated as the ratio of yeasts to fungi increased in the dual culture. A decrease in hyphal membrane potential and esterase activity was substantiated by the combined stains of DiBAC4(5) and CDFA-AM. Reduced metabolic function in conjunction with cell wall damage of A. flavus hindered the growth and biomass production of this fungus. Viability stains such as FUN-1 and DiBAC4(5) with CDFA-AM may assist in elucidating the biocontrol mechanism by allowing for the visualization of the antagonistic effect of yeast species on target fungi in situ, as well as for screening potent biocontrol yeast agents against fungal pathogens.

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References

  1. Hickey PC, Swift SR, Roca MG, Read ND. Live-cell imaging of filamentous fungi using vital fluorescent dyes and confocal microscopy. Method Microbiol. 2004;34:63–87.

    Article  Google Scholar 

  2. Lloyd D, Hayes AJ. Vigour, vitality and viability of microorganisms. FEMS Microbiol Lett. 1995;133:1–7.

    Article  CAS  Google Scholar 

  3. Millard PJ, Roth BL, Thai HP, Yu ST, Haugland RP. Development of the FUN-1 family of fluorescent probes for vacuole labeling and viability testing of yeasts. Appl Environ Microbiol. 1997;63:2897–905.

    CAS  PubMed  Google Scholar 

  4. Lass-Flörl C, Nagl M, Speth C, Ulmer H, Dierich MP, Würzner R. Studies of in vitro activities of Voriconazole and Itraconazole against Aspergillus hyphae using viability staining. Antimicrob Agents Chemother. 2001;45:124–8.

    Article  PubMed  Google Scholar 

  5. Marr KA, Koudadoust M, Black M, Balajee SA. Early events in macrophage killing of Aspergillus fumigatus conidia: new flow cytometric viability assay. Clin Diag Lab Immunol. 2001;8:1240–7.

    CAS  Google Scholar 

  6. Balajee SA, Marr KA. Conidial viability assay for rapid susceptibility testing of Aspergillus species. J Clin Microbiol. 2002;40:2741–5.

    Article  CAS  PubMed  Google Scholar 

  7. Bräuner T, Hülser DF, Strasser RJ. Comparative measurement of membrane potentials with microelectrodes and voltage-sensitive dyes. Biochimi Biophys Acta. 1984;771:208–16.

    Article  Google Scholar 

  8. Janisiewicz W, Korsten L. Biological control of postharvest diseases of fruits. Ann Rev Phytopathol. 2002;40:411–41.

    Article  CAS  Google Scholar 

  9. Spadaro D, Gullino ML. State of the art and future prospects of the biological control of postharvest fruit diseases. Int J Food Microbiol. 2004;91:185–94.

    Article  PubMed  Google Scholar 

  10. Jijakli MH, Lepoivre P. Characterization of an exo-β-1, 3 glucanase produced by Pichia anomala K, an antagonist of Botrytis cinerea on apples. Phytopathology. 1998;88:335–43.

    Article  CAS  PubMed  Google Scholar 

  11. Petersson S, Schnurer J. Biocontrol of mold growth in high-moisture wheat stored under airtight conditions by Pichia anomala, Pichia guillermondii, and Saccharomyces cerevisiae. Appl Environ Microbiol. 1995;61:1027–31.

    CAS  PubMed  Google Scholar 

  12. Petersson S, Scnurer J. Pichia anomala as a biocontrol agent of Penicillium roqueforti in high-moisture wheat, rye, barley, and oats stored under airtight conditions. Can J Microbiol. 1998;44:471–6.

    Article  CAS  Google Scholar 

  13. Hua SST, Baker JL, Flores-Espiritu M. Interactions of saprophytic yeasts with a nor mutant of Aspergillus flavus. Appl Environ Microbiol. 1999;65:2738–40.

    CAS  PubMed  Google Scholar 

  14. Hua SST. Progress in prevention of aflatoxin contamination in food by preharvest application of Pichia anomala WRL-076. In: Mendez-Vilas A, editor. Recent advances in multidisciplinary applied microbiology. Weinheim: Wiley-VCH Verlag GmbH&Co. KGaA; 2006. p. 322–6.

    Chapter  Google Scholar 

  15. Scudamore KA. Aspergillus toxin in food and animal feeding stuffs. In: Powell KA, Renwick A, Peberdy JF, editors. The genus Aspergillus. New York: Plenum Press; 1994. p. 59–71.

    Google Scholar 

  16. Commission of the European Community. Commission Directive 98/53/EC of July 1998 laying down the sampling methods and the methods of analysis for the official control of the levels of certain contaminants in food stuffs. Off Eur Communities Legislation. 1998; L201:93–101.

  17. U. S. Food and Drug Administration. Compliance policy guides manual. Washington, DC: U. S. FDA; 1996. Sec. 555.400, 268; Sec. 570.500, 299.

    Google Scholar 

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Acknowledgments

We thank Thomas McKeon. Glenn Barley and Bruce C. Campbell for their comments and reviews of the manuscript.

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Authors and Affiliations

  1. U. S. Department of Agriculture, Agricultural Research Service, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA

    Sui Sheng T. Hua, Maria T. Brandl, Bradley Hernlem, Jeffrey G. Eng & Siov Bouy L. Sarreal

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  1. Sui Sheng T. Hua
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  2. Maria T. Brandl
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  3. Bradley Hernlem
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  4. Jeffrey G. Eng
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  5. Siov Bouy L. Sarreal
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Correspondence to Sui Sheng T. Hua.

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Hua, S.S.T., Brandl, M.T., Hernlem, B. et al. Fluorescent Viability Stains to Probe the Metabolic Status of Aflatoxigenic Fungus in Dual Culture of Aspergillus flavus and Pichia anomala . Mycopathologia 171, 133–138 (2011). https://doi.org/10.1007/s11046-010-9352-z

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  • DOI: https://doi.org/10.1007/s11046-010-9352-z

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