Skip to main content
SpringerLink
Log in

Two rapid assays for screening of patulin biodegradation

  • Original Paper
  • Published:
International Journal of Environmental Science and Technology Aims and scope Submit manuscript

Abstract

The mycotoxin patulin is produced by the blue mould pathogen Penicillium expansum in rotting apples during postharvest storage. Patulin is toxic to a wide range of organisms, including humans, animals, fungi and bacteria. Wash water from apple packing and processing houses often harbours patulin and fungal spores, which can contaminate the environment. Ubiquitous epiphytic yeasts, such as Rhodosporidium kratochvilovae strain LS11 which is a biocontrol agent of P. expansum in apples, have the capacity to resist the toxicity of patulin and to biodegrade it. Two non-toxic products are formed. One is desoxypatulinic acid. The aim of the work was to develop rapid, high-throughput bioassays for monitoring patulin degradation in multiple samples. Escherichia coli was highly sensitive to patulin, but insensitive to desoxypatulinic acid. This was utilized to develop a detection test for patulin, replacing time-consuming thin layer chromatography or high-performance liquid chromatography. Two assays for patulin degradation were developed, one in liquid medium and the other in semi-solid medium. Both assays allow the contemporary screening of a large number of samples. The liquid medium assay utilizes 96-well microtiter plates and was optimized for using a minimum of patulin. The semi-solid medium assay has the added advantage of slowing down the biodegradation, which allows the study and isolation of transient degradation products. The two assays are complementary and have several areas of utilization, from screening a bank of microorganisms for biodegradation ability to the study of biodegradation pathways.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Notes

  1. PBS, Phosphate buffered saline.

References

  • Acar J, Gökmen V, Taydas EE (1998) The effects of processing technology on the patulin content of juice during commercial apple juice concentrate production. Z Lebensm Unters Forsch A 207:328–331

    Article  CAS  Google Scholar 

  • Anderson MS, Dutton MF, Harding K (1979) Production and degradation of patulin by Paecilomyces species, a common contaminant of silage. J Sci Food Agric 30:229–232

    Article  CAS  Google Scholar 

  • Boudergue C, Burel C, Dragacci S, Favrot MC, Fremy JM, Massimi C, Prigent P, Debongnie P, Pussemier L, Boudra H, Morgavi D, Oswald I, Perez A, Avantaggiato G (2009) Review of mycotoxin-detoxifying agents used as feed additives: mode of action, efficacy and feed/food safety. Scientific Report submitted to EFSA. 192 p. http://www.efsa.europa.eu/en/supporting/pub/22e.htm

  • Castoria R, Logrieco A (2008) Mycotoxins in fruits and major fruit-derived products—an overview. In: Ray RC, Ward OP (eds) Microbial biotechnology in horticulture, vol II., Science PublishersNew Hampshire, USA, pp 305–344

    Google Scholar 

  • Castoria R, De Curtis F, Lima G, Caputo L, Pacifico S, De Cicco V (2001) Aureobasidium pullulans (LS-30) an antagonist of postharvest pathogens of fruits: study on its modes of action. Postharvest Biol Technol 22:7–17

    Article  Google Scholar 

  • Castoria R, Caputo L, De Curtis F, De Cicco V (2003) Resistance of postharvest biocontrol yeasts to oxidative stress: a possible new mechanism of action. Phytopathology 93:564–572

    Article  CAS  Google Scholar 

  • Castoria R, Morena V, Caputo L, Panfili G, De Curtis F, De Cicco V (2005) Effect of the biocontrol yeast Rhodotorula glutinis strain LS11 on patulin accumulation in stored apples. Phytopathology 95:1271–1278

    Article  CAS  Google Scholar 

  • Castoria R, Mannina L, Durán Patrón R, Maffei F, Sobolev AP, De Felice DV, Pinedo-Rivilla C, Ritieni A, Ferracane R, Wright SAI (2011) Conversion of the mycotoxin patulin to the less toxic desoxypatulinic acid by the biocontrol yeast Rhodosporidium kratochvilovae strain LS11. J Agric Food Chem 59:11571–11578

    Article  CAS  Google Scholar 

  • Ciegler A (1978) Fungi that produce mycotoxins: conditions and occurrence. Mycopathologia 65:5–11

    Article  CAS  Google Scholar 

  • Ciegler A, Detroy RW, Lillehoj EB (1971) Patulin, penicillic acid, and other carcinogenic lactones. In: Ciegler A, Kadis S, Aji SJ (eds) Microbial toxins, vol 6. Academic Press Inc, New York, pp 409–434

    Google Scholar 

  • Commission Regulation (EC) No. 1881/2006, setting maximum levels for certain contaminants in foodstuffs. Off J Eur Union L364, 5

  • Commission Regulation (EC) No. 455/2004, amending Regulation (EC) No. 466/2001, as regards patulin. Off J Eur Union L74, 11

  • De Curtis F (1998) I lieviti nella lotta biologica contro patogeni fungini degli ortofrutticoli in postracolta, attività e meccanismi d’azione coinvolti. Ph.D. Dissertation, International Library of Florence and Rome, Italy

  • European Bioinformatics Institute website. https://www.ebi.ac.uk/chembldb/index.php/compound/inspect/CHEMBL294018

  • Fliege R, Metzler M (2000) Electrophilic properties of patulin. N-acetylcysteine and glutathione adducts. Chem Res Toxicol 13:373–381

    Article  CAS  Google Scholar 

  • Gonçalves AB, Paterson RRM, Lima N (2006) Survey and significance of filamentous fungi from tap water. Int J Hyg Environ-Health 209:257–264

    Article  Google Scholar 

  • Gromadzka K, Waskiewicz A, Golinski P, Swietlik J (2009) Occurrence of estrogenic mycotoxin—Zearalenone in aqueous environmental samples with various NOM content. Water Res 43:1051–1059

    Article  CAS  Google Scholar 

  • Hageskal G, Knutsen AK, Gaustad P, Sybren de Hoog G, Skaar I (2006) Diversity and Significance of Mold Species in Norwegian Drinking Water. Appl Environ Microbiol 72(12):7586–7593

    Article  CAS  Google Scholar 

  • Hageskal G, Gaustad P, Heier BT, Skaar I (2007) Occurrence of moulds in drinking water. J Appl Microbiol 102:774–780

    Article  CAS  Google Scholar 

  • Halász A, Lásztity R, Abonyi T, Bata A (2009) Decontamination of mycotoxin-containing food and feed by biodegradation. Food Rev Int 25:284–298

    Article  Google Scholar 

  • Harwig J, Scott PM, Kennedy BPC, Chen YK (1973) Disappearance of patulin from apple juice fermented by Saccharomyces spp. J Inst Can Sci Technol Aliment 6:45–46

    Article  CAS  Google Scholar 

  • He J, Zhou T, Young JC, Boland GJ, Scott PM (2010) Chemical and biological transformations for detoxification of trichothecene mycotoxins in human and animal food chains: a review. Trends Food Sci Technol 21:67–76

    Article  CAS  Google Scholar 

  • Hoerger CC, Schenzel J, Strobel BW, Bucheli TD (2009) Analysis of selected phytotoxins and mycotoxins in environmental samples. Anal Bioanal Chem 395:1261–1289

    Google Scholar 

  • Karlovsky P (1999) Biological detoxification of fungal toxins and its use in plant breeding, feed and food production. Nat Toxins 7:1–23

    Article  CAS  Google Scholar 

  • Lilly VG, Barnett HL (1951) Physiology of the fungi. McGraw-Hill, New York

    Google Scholar 

  • Lima G, De Curtis F, Castoria R, De Cicco V (2003) Integrated control of apple postharvest pathogens and survival of biocontrol yeasts in semi-commercial conditions. Eur J Plant Pathol 109:341–349

    Article  CAS  Google Scholar 

  • Lima G, Castoria R, De Curtis F, Raiola A, Ritieni A, De Cicco V (2011) Integrated control of blue mould using new fungicides and biocontrol yeasts lowers levels of fungicide residues and patulin contamination in apples. Postharvest Biol Technol 60:164–172

    Google Scholar 

  • MacDonald S, Long M, Gilbert J (2000) Liquid chromatographic method for determination of patulin in clear and cloudy apple juices and apple puree: collaborative study. J AOAC Int 83:1387–1394

    CAS  Google Scholar 

  • McKinley ER, Carlton WW (1991) Patulin. In: Sharma RP, Salunkhe DK (eds) Mycotoxins and Phytoalexins. CRC Press, Boca Raton, pp 191–236

    Google Scholar 

  • Moake MM, Padilla-Zakour OI, Worobo RW (2005) Comprehensive review of patulin control methods in foods. Compr Rev Food Sci Food Saf 1:8–21

    Article  Google Scholar 

  • Moss MO, Long MT (2002) Fate of patulin in the presence of yeast Saccharomyces cerevisiae. Food Addit Contam 19:387–399

    Article  CAS  Google Scholar 

  • Pereira VJ, Basílio MC, Fernandes D, Domingues M, Paiva JM, Benoliel MJ, Crespo MT, San Romão MV (2009) Occurrence of filamentous fungi and yeasts in three different drinking water systems. Water Res 43:3813–3819

    Article  CAS  Google Scholar 

  • Pérez-Guerra N, Torrado-Agrasar A, López-Macias C, Pastrana L (2003) Main characteristics and applications of solid substrate fermentation. Electron J Environ Agric Food Chem 2(3):343–350

    Google Scholar 

  • Reiss J (1975) Bacillus subtilis; a sensitive bioassay for patulin. Bull Environ Contam Toxicol 13(6):689–691

    Article  CAS  Google Scholar 

  • Ricelli A, Baruzzi F, Solfrizzo M, Morea M, Fanizzi FP (2007) Biotransformation of patulin by Gluconobacter oxydans. Appl Environ Microbiol 73(3):785–792

    Article  CAS  Google Scholar 

  • Robinson T, Singh D, Nigam P (2001) Solid-state fermentation: a promising microbial technology for secondary metabolite production. Appl Microbiol Biotechnol 55:284–289

    Article  CAS  Google Scholar 

  • Sammon NB, Harrower KM, Fabbro LD, Reed RH (2010) Incidence and distribution of microfungi in a treated municipal water supply system in sub-tropical Australia. Int J Environ Res Public Health 7:1597–1611

    Google Scholar 

  • Sammon NB, Harrower KM, Fabbro LD, Reed RH (2011) Three potential sources of microfungi in a treated municipal water supply system in sub-tropical Australia. Int J Environ Res Public Health 8:713–732

    Article  Google Scholar 

  • Scott PM, Kennedy B, Van Walbeek W (1972) Desoxypatulinic acid from a patulin-producing strain of Penicillium patulum. Experientia 28:1252

    Article  CAS  Google Scholar 

  • Siqueira VM, Oliveira HMB, Santos C, Patterson RRM, Gusmão NB, Lima N (2011) Filamentous fungi in drinking water, particularly in relation to biofilm formation. Int J Environ Res Public Health 8:456–469

    Article  CAS  Google Scholar 

  • Steiman R, Seigle-Murandi F, Sage L, Krivobok S (1989) Production of patulin by Micromycetes. Mycopathologia 105:129–133

    Article  CAS  Google Scholar 

  • Stinson EE, Osman SF, Huhtanen CN, Bills DD (1978) Disappearance of patulin during alcoholic fermentation of apple juice. Appl Environ Microbiol 36(4):620–622

    CAS  Google Scholar 

  • Sumbu ZL, Thonart P, Bechet J (1983) Action of patulin on a yeast. Appl Environ Microbiol 45(1):110–115

    CAS  Google Scholar 

  • US EPA Estimation Program Interface (EPI) Suite. Ver.4.0. Jan, 2009. http://www.epa.gov/oppt/exposure/pubs/episuite.htm

  • Vansteelandt M, Kerzaon I, Blanchet E, Fossi Tankoua O, Robiou Du, Pont T, Joubert Y, Monteau F, Le Bizec B, Pouchus YF, Grovel O (2012) Patulin and secondary metabolite production by marine-derived Penicillium strains. Fungal Biol 116(9):954–961

    Article  CAS  Google Scholar 

  • Wright SAI, Zumoff CH, Schneider L, Beer SV (2001) Strain Eh318 of Pantoea agglomerans produces two antibiotics that inhibit Erwinia amylovora in vitro. Appl Environ Microbiol 67(1):284–292

    Article  CAS  Google Scholar 

  • Wu QK, Jezkova A, Yuan Z, Pavlikova L, Dohnal V, Kuca K (2009) Biological degradation of aflatoxins. Drug Metab Rev 41:1–7

    Article  Google Scholar 

  • Yates IE (1986) Bioassay systems and their use in the diagnosis of mycotoxicoses. In: Richard JL, Thurston JR (eds) Diagnosis of mycotoxicoses. Martinus Nijhoff, The Hague, pp 333–378

    Chapter  Google Scholar 

Download references

Acknowledgments

We would like to express our sincere thanks to Dr. Anders Falk, who performed the statistical analysis. We are truly grateful to student Paolo D’Apruzzo, who participated in the development of the filter assay. We would like also to acknowledge Dr. Alison Hill, University of Exeter, United Kingdom, for her contributions to the discussion, to Fig. 1 and for the critical reading of the manuscript. We are indebted to Dr. Rosa Durán-Patrón, University of Cádiz, Spain, who enabled C.P.-R. to participate in this work. Dr. Duran-Patrón has been collaborating on the structural elucidation of the biodegradation products of patulin through a bilateral project entitled: “Studio del pathway di biodegradazione della micotossina patulina operata da un lievito basidiomicete”, with funding from the Ministero dell’Istruzione, dell’Università e della Ricerca (project number IT088MB951) in Italy, and by the Ministerio de Ciencia e Innovación (project number HI2007-0026) in Spain. The research was overall funded by the Italian Ministry for University and Scientific Research (MIUR), through the two projects: PRIN n. 2006072204 and “Incentivazione alla mobilità di studiosi stranieri e italiani residenti all’estero”, (DM 1.2.2005, n.18).

Author information

Author notes

  1. S. A. I. Wright

    Present address: Department of Electronics, Mathematics and Natural Sciences, University of Gävle, SE-801 76 Gävle, Gävle, Sweden

Authors and Affiliations

  1. Department of Agricultural, Environmental and Food Sciences, Università degli Studi del Molise, Via Francesco De Sanctis, 86100, Campobasso, Italy

    S. A. I. Wright, D. V. de Felice, G. Ianiri, F. De Curtis & R. Castoria

  2. Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Cádiz, Campus Universitario Puerto Real, 11510, Puerto Real, Cádiz, Spain

    C. Pinedo-Rivilla

Authors
  1. S. A. I. Wright
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. V. de Felice
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Ianiri
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C. Pinedo-Rivilla
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. F. De Curtis
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R. Castoria
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to S. A. I. Wright or R. Castoria.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wright, S.A.I., de Felice, D.V., Ianiri, G. et al. Two rapid assays for screening of patulin biodegradation. Int. J. Environ. Sci. Technol. 11, 1387–1398 (2014). https://doi.org/10.1007/s13762-013-0325-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13762-013-0325-x

Keywords

Search

Navigation