Mycotoxin Research

, Volume 34, Issue 2, pp 91–97 | Cite as

Multimycotoxin analysis of South African Aspergillus clavatus isolates

  • C. J. Botha
  • M. Truter
  • M. Sulyok
Original Article


Aspergillus clavatus poisoning is a neuromycotoxicosis of ruminants that occurs sporadically across the world after ingestion of infected feedstuffs. Although various toxic metabolites are synthesized by the fungus, it is not clear which specific or group of mycotoxins induces the syndrome. A. clavatus isolates were deposited in the culture collection of the Biosystematics Division, Plant Protection Research Institute, Agricultural Research Council during incidences of livestock poisoning (1988–2016). Six isolates were still viable and these plus three other South African isolates that were also previously deposited in the collection were positively identified as A. clavatus based on morphology and ß-tubulin sequence data. The cultures were screened for multiple mycotoxins using a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method. Twelve A. clavatus metabolites were detected. The concentrations of the tremorgenic mycotoxins (i.e., tryptoquivaline A and its related metabolites deoxytryptoquivaline A and deoxynortryptoquivaline) were higher than patulin and cytochalasin E. Livestock owners should not feed A. clavatus-infected material to ruminants as all the South African A. clavatus isolates synthesized the same compounds when cultured under similar conditions.


Aspergillus clavatus Cytochalasin E Patulin Tremorgenic Tryptoquivaline 



The authors would also like to express their gratitude to Marie Smith, Stats4science, who performed the statistical analysis and Danielle Henn who assisted with the drawing of the chemical structures.

Funding information

This work is based on research supported in part by the National Research Foundation of South Africa (Grant number 103747).

Compliance with ethical standards

Conflict of interest



  1. Austin WL, Wind M, Brown KS (1982) Differences in the toxicity and teratogenicity of cytochalasin D and E in various mouse strains. Teratology 25(1):11–18. CrossRefPubMedGoogle Scholar
  2. Botha CJ, Legg MJ, Truter M, Sulyok M (2014) Multitoxin analysis of Aspergillus clavatus infected feed samples implicated in two outbreaks of neuromycotoxicosis in cattle in South Africa. Onderstepoort J Vet Res 81(1):6.
  3. Büchi G, Kitaura Y, Yuan SS, Wright HE, Clardy J, Demain AL, Glinsukon T, Hunt N, Wogan GN (1973) Structure of cytochalasin E, a toxic metabolite of Aspergillus clavatus. J Am Chem Soc 95(16):5423–5425. CrossRefPubMedGoogle Scholar
  4. Büchi G, Luk KC, Kobbe B, Townsend JM (1977) Four new mycotoxins of Aspergillus clavatus related to tryptoquivaline. J Org Chem 42(2):244–246. CrossRefPubMedGoogle Scholar
  5. Clardy J, Springer JP, Koenig T, Büchi G, Matsuo K, Wightman R (1975) Tryptoquivaline and tryptoquivalone, two tremorgenic metabolites of Aspergillus clavatus. J Am Chem Soc 97(3):663–665. CrossRefPubMedGoogle Scholar
  6. Gilmour JS, Inglis DM, Robb J, Maclean M (1989) A fodder mycotoxicosis of ruminants caused by contamination of a distillery by-product with Aspergillus clavatus. Vet Rec 124(6):133–135. CrossRefPubMedGoogle Scholar
  7. Glass NL, Donaldson GC (1995) Development of premier sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl Environ Microbiol 61:1323–1330PubMedPubMedCentralGoogle Scholar
  8. Glinsukon T, Yuan SS, Wightman R, Kitaura Y, Büchi G, Shank RC, Wogan GN, Christensen CM (1974) Isolation and purification of cytochalasin E and two tremorgens from Aspergillus clavatus. Plant Foods Man 1:113–119CrossRefGoogle Scholar
  9. Hayes AW, Phillips TD, Williams WL (1978) Acute toxicity of patulin. Toxicol Appl Pharmacol 45:275–276Google Scholar
  10. Kellerman TS, Pienaar JG, Van der Westhuizen GCA, Anderson LAP, Naudé TW (1976) A highly fatal tremorgenic mycotoxicosis of cattle caused by Aspergillus clavatus. Onderstepoort J Vet 43:147–154Google Scholar
  11. Kellerman TS, Coetzer JAW, Naudé TW, Botha CJ (2005) Plant poisonings and mycotoxicoses of livestock in southern Africa, 2nd edn. Oxford University Press, Cape TownGoogle Scholar
  12. Klich MA (2002) Identification of common Aspergillus species. Centraalbureau voor Schimmelcultures, UtrechtGoogle Scholar
  13. Lopez-Diaz TM, Flannigan B (1997) Production of patulin and cytochalasin E by Aspergillus clavatus during malting of barley and wheat. Int J Food Microbiol 35(2):129–136. CrossRefPubMedGoogle Scholar
  14. 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. CrossRefPubMedGoogle Scholar
  15. 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 28:542–555Google Scholar
  16. McKenzie RA, Kelly MA, Shivas RG, Gibson JA, Cook PJ, Widderick K, Guilfoyle AF (2004) Aspergillus clavatus tremorgenic neurotoxicosis in cattle fed sprouted grains. Aust Vet J 82(10):635–638. CrossRefPubMedGoogle Scholar
  17. McKinley ER, Carlton WW (1979) Patulin mycotoxicosis in Swiss ICR mice. Food Cosmet Toxicol 18:181–187CrossRefGoogle Scholar
  18. Payne RW (2014) Introduction to GenStat for Windows, 17th edn. VSN International, Hemel HempsteadGoogle Scholar
  19. Riet-Correa F, Rivero R, Odriozola E, De Lourdes Adrien M, Medeiros RMT, Schild AL (2013) Mycoxicoses of ruminants and horses. J Vet Diagn Investig 25(6):692–708. CrossRefGoogle Scholar
  20. Sabater-Vilar M, Maas RFM, De Bosschere H, Ducatelle R, Fink-Gremmels J (2004) Patulin produced by an Aspergillus clavatus isolated from feed containing malting residues associated with a lethal neurotoxicosis in cattle. Mycopathologia 158(4):419–426. CrossRefPubMedGoogle Scholar
  21. Samson RA, Visagie CM, Houbraken J, Hong S-B, Hubka V, Klaassen CHW, Perrone G, Seifert KA, Susca A, Tanney JB, Varga J, Kocsubé S, Szigeti G, Yaguchi T, Frisvad JC (2014) Phylogeny, identification and nomenclature of the genus Aspergillus. Stud Mycol 78:141–173. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Schlosberg A, Zadikov I, Perl S, Yakobson B, Varod Y, Elad D, Rapoprt E, Handji V (1991) Aspergillus clavatus as the probable cause of a lethal mass neurotoxicosis in sheep. Mycopathologia 114(1):35–39. CrossRefGoogle Scholar
  23. Swofford DL (2002). PAUP*. phylogenetic analysis using parsimony (*and other methods). version 4.0b10. Sinauer Associates, Sunderland, MA, USA.Google Scholar
  24. Uhlig S, Botha CJ, Vrålstad T, Rolén E, Miles CO (2009) Indole–diterpenes and ergot alkaloids in Cynodon dactylon (Bermuda grass) infected with Claviceps cynodontis from an outbreak of tremors in cattle. J Agric Food Chem 57(23):11112–11119. CrossRefPubMedGoogle Scholar
  25. Varga J, Due M, Frisvad JC, Samson RA (2007) Taxonomic revision of Aspergillus section Clavati based on molecular, morphological and physiological data. Stud Mycol 59:89–106. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Society for Mycotoxin Research and Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Paraclinical Sciences, Faculty of Veterinary ScienceUniversity of PretoriaOnderstepoortSouth Africa
  2. 2.Biosystematics DivisionAgricultural Research Council - Vegetable and Ornamental PlantsPretoriaSouth Africa
  3. 3.Centre for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln)University of Natural Resources and Life SciencesTullnAustria

Personalised recommendations