Putative neuromycotoxicoses in an adult male following ingestion of moldy walnuts

C. J. Botha, C. M. Visagie, M. Sulyok

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.