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Putative neuromycotoxicoses in an adult male following ingestion of moldy walnuts

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Abstract

A tremorgenic syndrome occurs in dogs following ingestion of moldy walnuts, and Penicillium crustosum has been implicated as the offending fungus. This is the first report of suspected moldy walnut toxicosis in man. An adult male ingested approximately eight fungal-infected walnut kernels and after 12 h experienced tremors, generalized pain, incoordination, confusion, anxiety, and diaphoresis. Following symptomatic and supportive treatment at a local hospital, the man made an uneventful recovery. A batch of walnuts (approximately 20) was submitted for mycological culturing and identification as well as for mycotoxin analysis. Penicillium crustosum Thom was the most abundant fungus present on walnut samples, often occurring as monocultures on isolation plates. Identifications were confirmed with DNA sequences. The kernels and shells of the moldy walnuts as well as P. crustosum isolates plated on yeast extract sucrose (YES) and Czapek yeast autolysate (CYA) agars and incubated in the dark at 25 °C for 7 days were screened for tremorgenic mycotoxins and known P. crustosum metabolites using a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method. A relatively low penitrem A concentration of only 1.9 ng/g was detected on the walnut kernels when compared to roquefortine C concentrations of 21.7 μg/g. A similar result was obtained from P. crustosum isolates cultured on YES and CYA, with penitrem A concentrations much lower (0.6–6.4 μg per g mycelium/agar) compared to roquefortine C concentrations (172–1225 μg/g). The authors surmised that besides penitrem A, roquefortine C might also play an additive or synergistic role in intoxication of man.

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References

  • Arnold DL, Scott PM, McGuire PF, Harwig J, Nera EA (1978) Acute toxicity studies on roquefortine and PR toxin, metabolites of Penicillium roqueforti, in the mouse. Food Cosmet Toxicol 16:369–371

    Article  CAS  PubMed  Google Scholar 

  • Blakeslee AF (1915) Lindner’s roll tube method of separation cultures. Phytopathology 5:68–69

    Google Scholar 

  • Braselton WE, Rumler PC (1996) MS/MS screen for the tremorgenic mycotoxins roquefortine and penitrem A. J Vet Diagn Investig 8:515–518

    Article  CAS  Google Scholar 

  • Bünger J, Westphal G, Mönnich A, Hinnendahl B, Hallier E, Müller M (2004) Cytotoxicity of occupationally and environmentally relevant mycotoxins. Toxicology 202:199–211. https://doi.org/10.1016/j.tox.2004.05.007

    Article  CAS  PubMed  Google Scholar 

  • Ciegler A, Vesonder RF, Cole RJ (1976) Tremorgenic mycotoxins. In: Rodricks JV (ed) Mycotoxins and other related food problems, vol. 149. Adv Chem Ser, Am Chem Soc, Washington, pp 163–177

  • Cole RJ, Dorner JW, Cox RH, Raymond LW (1983) Two classes of alkaloid mycotoxins produced by Penicillium crustosum Thom isolated from contaminated beer. J Agric Food Chem 31(3):655–657

    Article  CAS  PubMed  Google Scholar 

  • De Jesus AE, Steyn PS, Van Heerden FR, Vleggaar R, Wessels PL (1983) Tremorgenic mycotoxins from Penicillium crustosum: isolation of penitrems A–F and the structure elucidation and absolute configuration of penitrem A. J Chem Soc Perkin Trans I:1847–1856

    Article  Google Scholar 

  • Eriksen GS, Hultin-Jäderlund K, Moldes-Anaya A, Schönheit J, Bernhoft A, Jaeger G, Rundberget T, Skaar I (2010) Poisoning of dogs with tremorgenic Penicillium toxins. Med Mycol 48:188–196. https://doi.org/10.3109/13693780903225821

    Article  CAS  PubMed  Google Scholar 

  • Frisvad JC, Samson RA (2004) Polyphasic taxonomy of Penicillium subgenus Penicillium. A guide to identification of food and air-borne terverticillate Penicillia and their mycotoxins. Stud Mycol 49:1–174

    Google Scholar 

  • Frisvad JC, Smedsgaard J, Larsen TO, Samson RA (2004) Mycotoxins, drugs and other extrolites produced by species in Penicillium subgenus Penicillium. Stud Mycol 49:201–241

    Google Scholar 

  • Glass NL, Donaldson GC (1995) Development of premier sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl Environ Microb 61:1323–1330

    CAS  Google Scholar 

  • Hayes AW, Presley DB, Neville JA (1976) Acute toxicity of penitrem A in dogs. Toxicol Appl Pharm 35:311–320

    Article  CAS  Google Scholar 

  • Kalinina SA, Jagels A, Hickert S, Mauriz Marques LM, Cramer B, Humpf H-U (2018) Detection of the cytotoxic penitrems A−F in cheese from the European single market by HPLC-MS/MS. J Agric Food Chem 66:1264–1269. https://doi.org/10.1021/acs.jafc.7b06001

    Article  CAS  PubMed  Google Scholar 

  • Kalyaanamoorthy S, Minh BQ, Wong TKF, von Haeseler A, Jermiin LS (2017) ModelFinder: fast model selection for accurate phylogenetic estimates. Nat Methods 14:587–589. https://doi.org/10.1038/nmeth.4285

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol 30:772–780. https://doi.org/10.1093/molbev/mst010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Knaus H-G, McManus OB, Lee SH, Schmalhofer WA, Garcia-Calvo M, Helms LMH, Sanchez M, Giangiacomo K, Reuben JP, Smith AB, Kaczorowski GJ, Garcia ML (1994) Tremorgenic indole alkaloids potently inhibit smooth muscle high conductance calcium activated potassium channels. Biochemistry 33:5819–5828

    Article  CAS  PubMed  Google Scholar 

  • Lamboni Y, Nielsen KF, Linnemann AR, Gezgin Y, Hell K, Nout MJR, Smid EJ, Tamo M, van Boekel MAJS, Hoof JB, Frisvad JC (2016) Diversity in secondary metabolites including mycotoxins from strains of Aspergillus section Nigri isolated from raw cashew nuts from Benin. West Africa Plos One 11(10):e0164310. https://doi.org/10.1371/journal.pone.0164310

    Article  CAS  PubMed  Google Scholar 

  • Lewis PR, Donoghue MB, Hocking AD, Cook L, Granger LV (2005) Tremor syndrome associated with fungal toxin: sequelae of food contamination. Med J Aust 182(11):582–584

    PubMed  Google Scholar 

  • Lowes NR, Smith RA, Beck BE (1992) Roquefortine in the stomach contents of dogs suspected of strychnine poisoning in Alberta. Can Vet J 33:535–538

    CAS  PubMed  PubMed Central  Google Scholar 

  • Malachova A, Sulyok M, Beltran E, Berthiller F, Krska R (2014) Optimization and validation of a quantitative liquid chromatography-tandem mass spectrometric method covering 295 bacterial and fungal metabolites including all regulated mycotoxins in four model food matrices. J Chromatogr A 1362:145–156. https://doi.org/10.1016/j.chroma.2014.08.037

    Article  CAS  PubMed  Google Scholar 

  • Malachova A, Sulyok M, Beltran E, Berthiller F, Krska R (2015) Multi-toxin determination in food - the power of “dilute and shoot” approaches in LC-MS-MS. LC GC Eur 28:542–555

    Google Scholar 

  • Moldes-Anaya A, Fonnum F, Eriksen GS, Rundberget T, Walaas SI, Wigestrand MB (2011) In vitro neuropharmacological evaluation of penitrem-induced tremorgenic syndromes: importance of the GABAergic system. Neurochem Int 59:1074–1081. https://doi.org/10.1016/j.neuint.2011.08.014

    Article  CAS  PubMed  Google Scholar 

  • Moldes-Anaya A, Rundberget T, Faeste CK, Eriksen GS, Bernhoft A (2012) Neurotoxicity of Penicillium crustosum secondary metabolites: tremorgenic activity of orally administered penitrem A and thomitrem A and E in mice. Toxicon 60:1428–1435. https://doi.org/10.1016/j.toxicon.2012.10.007

    Article  CAS  PubMed  Google Scholar 

  • Munday JS, Thompson D, Finch SC, Babu JV, Wilkins AL, di Menna ME, Miles CO (2008) Presumptive tremorgenic mycotoxicosis in a dog in New Zealand, after eating mouldy walnuts. New Zeal Vet J 56(3):145–147. https://doi.org/10.1080/00480169.2008.36823

    Article  CAS  Google Scholar 

  • Naudé TW, O’Brien OM, Rundberget T, McGregor ADG, Roux C, Flåøyen A (2002) Tremorgenic neuromycotoxicosis in 2 dogs ascribed to the ingestion of penitrem A and possibly roquefortine in rice contaminated with Penicillium crustosum. J S Afr Vet Assoc 73:211–215. https://doi.org/10.4102/jsava.v73i4.589

    Article  PubMed  Google Scholar 

  • Nguyen LT, Schmidt HA, Von Haeseler A, Minh BQ (2015) IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Mol Biol Evol 32:268–274. https://doi.org/10.1093/molbev/msu300

    Article  CAS  PubMed  Google Scholar 

  • Norris PJ, Smith CCT, De Belleroche J, Bradford HF, Mantle PG, Thomas AJ, Penny RHC (1980) Actions of tremorgenic fungal toxins on neurotransmitter release. J Neurochem 34:33–42

    Article  CAS  PubMed  Google Scholar 

  • Puls R, Ladyman E (1988) Roquefortine toxicity in a dog. Can Vet J 29:569

    CAS  PubMed  PubMed Central  Google Scholar 

  • Richard JL, Bacchetti P, Arp LH (1981) Moldy walnut toxicosis in a dog, caused by the mycotoxin, penitrem A. Mycopathologia 76:55–58

    Article  CAS  PubMed  Google Scholar 

  • Rundberget T, Skaar I, O’Brien O, Flåøyen A (2004) Penitrem and thomitrem formation by Penicillium crustosum. Mycopathologia 157:349–357

    Article  PubMed  Google Scholar 

  • Tiwary AK, Puschner B, Poppenga RH (2009) Using roquefortine C as a biomarker for penitrem A intoxication. J Vet Diagn Investig 21:237–239

    Article  Google Scholar 

  • Visagie CM, Houbraken J, Frisvad JC, Hong SB, Klaassen CHW, Perrone G, Seifert KA, Varga J, Yaguchi T, Samson RA (2014) Identification and nomenclature of the genus Penicillium. Stud Mycol 78:343–371. https://doi.org/10.1016/j.simyco.2014.09.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Young KL, Villar D, Carson TL, Imerman PM, Moore RA, Bottoff MR (2003) Tremorgenic mycotoxin intoxication with penitrem A and roquefortine in two dogs. J Am Vet Med Assoc 222:52–53

    Article  PubMed  Google Scholar 

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Acknowledgements

This work is based on research supported in part by the National Research Foundation of South Africa (grant number 103747). The authors would like to thank Dr. Leopold Podlashuc for bringing this case to our attention, and we would also like to express our gratitude to Danielle Henn who assisted with the drawing of the chemical structures.

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Correspondence to C. J. Botha.

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Botha, C.J., Visagie, C.M. & Sulyok, M. Putative neuromycotoxicoses in an adult male following ingestion of moldy walnuts. Mycotoxin Res 35, 9–16 (2019). https://doi.org/10.1007/s12550-018-0326-1

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