DOI: 10.1002/chem.

201406655 Correspondence

& Natural Products

Comment on “Chemical and Toxicological Investigations of

a Previously Unknown Poisonous European Mushroom Tricholoma
terreum”
Paolo Davoli,*[a] Marco Floriani,[b] Francesca Assisi,[c] Karl Kob,[d] and Nicola Sitta[e]

Abstract: Recent findings casting doubts over the edibility considerations. Accordingly, T. terreum should remain listed
of the European mushroom Tricholoma terreum are ques- among edible mushroom species.
tioned on the basis of mycological and mycotoxicological

Tricholoma terreum (Schaeff.) P. Kumm. (Tricholomataceae,
Agaricales, Basidiomycotina) is a well-known fungal species
producing small to medium-sized, agaricoid fruiting bodies
that are characterised by a greyish, typically felty cap, white to
light greyish gills, white spore print, weak smell, scanty or no
velar remnants and non-yellowing context.[1] It is widespread
and common in most European countries, usually growing on
sandy or loamy soils, mainly in association with conifers.
In mushroom field guides published throughout Europe T.
terreum is considered an edible species[3] and in the FAO’s
compendium of wild edible mushrooms it is listed as “edible”
or “food”.[4] In Europe, sporophores of T. terreum and allied spe-
cies, such as Tricholoma scalpturatum (Fr.) Qul., Tricholoma or-
irubens Qul. and others, are collected and safely eaten in Italy,
France, Germany, Spain and elsewhere; the existence of several
vernacular names in different languages—the most known and
used are moretta in Italian, petit-gris in French and negrilla in
Spanish—further attests the popular knowledge and the long
tradition of harvesting of these species for food purposes. Ac-
cordingly, T. terreum is included in the positive lists of marketa-
ble mushroom species for Italy, Spain, Belgium and Germany.[5]
Also mycology texts and field guides for countries outside
Europe, for example, United States,[6a] Japan[6b] and China,[6c, d]
confirm the edibility of T. terreum, although it is not certain
whether the species identified as such in North America and
Asia is the same as collected in Europe.
Most recently, a group of Chinese researchers led by Ji-Kai
Liu has reported in this journal the isolation and structure elu-
cidation of an array of novel fungal metabolites from sporo-
phores of T. terreum collected in France.[7] Purification of non-
polar fractions of T. terreum extracts led to the isolation of fif-
teen new triterpenoids, namely terreolides A–F and saponaceo-
lides H–P,[7] along with known saponaceolide B.[8] Saponaceo-
lide B and M (Figure 1) displayed toxicity in mice with LD50

[a] Dr. P. Davoli

via Pellegrini 2/18
41058 Vignola MO (Italy)
E-mail: paolo-davoli@libero.it
[b] M. Floriani
via dei Caldonazzi 44 - Serso Figure 1. Structures of saponaceolides B and M from Tricholoma terreum.
38057 Pergine Valsugana TN (Italy)
[c] Dr. F. Assisi
S.C. Centro Antiveleni, Dipartimento Emergenza Urgenza - E.A.S.
values of 88.3 and 63.7 mg kg 1, respectively; in particular, oral
Azienda Ospedaliera Niguarda Ca’ Granda
piazza Ospedale Maggiore 3 administration with saponaceolides B and M resulted in the in-
20162 Milano (Italy) crease of serum creatine kinase (CK) levels in mice. Saponaceo-
[d] Dr. K. Kob lide B and M were found to occur in T. terreum fruiting bodies
via Montello 13/9 at levels of 414 and 184 mg kg 1 dry weight, respectively. On
39100 Bolzano (Italy)
the basis of these findings, Yin et al. concluded that sapona-
[e] N. Sitta
ceolide B and M are responsible for the toxicity of T. terreum
Loc. Farn 32
40042 Lizzano in Belvedere BO (Italy) and, therefore, they referred to this fungal species as a ‘hitherto
Supporting information for this article is available on the WWW under unknown poisonous European mushroom’.[7] Moreover, given
http://dx.doi.org/10.1002/chem.201406655. that the increase in serum CK levels has always been observed

Chem. Eur. J. 2016, 22, 1 – 4 1  2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim & &

These are not the final page numbers! ÞÞ


in cases of mushroom-induced rhabdomyolysis, such as those
well described in the literature for Tricholoma equestre (L.) P.
Kumm.,[9, 10] Yin and colleagues also concluded that the out-
breaks of rhabdomyolysis which have been reported from
France and Poland after consumption of high amounts of T.
equestre in consecutive meals might be due to T. terreum in-
stead, as T. terreum might have been misidentified for T. eque-
stre, ‘given that the two mushrooms grow in similar environ-
ments and that they belong to the same genus’.[7] In the course
of their study, Yin et al. also investigated sporophores of T.
equestre from the very same region in France where outbreaks
of mushroom-induced rhabdomyolysis had originally occurred
after consumption of T. equestre. By contrast to T. terreum,
whose nonpolar fractions displayed toxicity in mice, polar frac-
tions from extracts of T. equestre were found toxic to mice and
induced increase in CK levels; neither saponaceolide B or M
were detected in T. equestre. On the basis of these findings, Yin
et al. concluded that T. equestre and T. terreum must contain
different toxins, but attempts to isolate the compounds re-
sponsible for such toxicity in T. equestre failed.[7]
From the viewpoint of natural product chemistry, the paper
by Yin et al.[7] is scientifically sound and certainly adds to our
knowledge of bioactive fungal metabolites of the genus Tricho-
loma. Unfortunately, however, the final speculations clash
heavily with well accepted mycological and mycotoxicological
data. The conclusions that Liu and his team have drawn are
misleading from a mycotoxicological perspective, as they cast
doubts on the edibility of a mushroom species that has been
always recognized as safe so far. To the best of our knowledge,
in fact, there are no documented cases of poisoning due to
the ingestion of T. terreum in Europe and elsewhere, and this
species has never been reported in the medical literature to
induce any symptom related to rhabdomyolysis after con-
sumption.
At the outset it should be pointed out that despite belong-
ing to the same genus, T. terreum and T. equestre are morpho-
logically well distinct species and their overall colours are suffi-
cient to tell them apart in the field.[1, 3b, 11] It is therefore awk-
ward to assume that the victims of poisonings attributed to T.
equestre, who had been collecting the latter for years, might
have confused it with the ‘allegedly’ toxic T. terreum. T. eque-
stre-induced rhabdomyolysis is well described in the literature,
both in humans and in mice, following ingestion in consecu-
tive meals.[12] Interestingly, and most intriguingly, myotoxic ef-
fects were also observed in mice after administration of other
edible wild and cultivated mushroom species.[12b, d] More re-
cently, a human case of mushroom-induced rhabdomyolysis
has been reported following consumption of Boletus edulis s.l.
and Leccinum spp.,[13a] a group of commercially relevant wild
mushroom species with a long tradition of consumption
worldwide,[14] and another case has been also described in the
medical literature after ingestion of cultivated Agaricus bisporus
(J.E. Lange) Imbach.[13b] Hence, apparently there is growing evi-
dence suggesting that rhabdomyolysis following the ingestion
of a large amount of edible mushrooms in consecutive meals
can be a symptom not or weakly related to the fungal species
ingested. Whether such myotoxicity following prolonged con-
& & Chem. Eur. J. 2016, 22, 1 – 4 www.chemeurj.org
Correspondence
sumption of mushrooms in relevant quantity has any bearing
with specific toxic compounds remains unanswered at present,
and certainly warrants further investigations.
With regards to the toxicity of T. terreum extracts observed
by Yin et al., we cannot help noticing that crude extracts of T.
terreum and fractions thereof, including purified saponaceolide
B and M, were fed orally to mice without any prior treatment.[7]
However, traditionally T. terreum is never eaten raw, but it is
always consumed after cooking. In our opinion, Liu and his
team should have also verified experimentally whether sapona-
ceolide B and M are capable of withstanding prolonged heat-
ing at boiling temperatures such as those which are typically
encountered during cooking. In the absence of such a proof, it
is tempting to speculate that lipophilic saponaceolide B and M
might be somewhat destroyed/inactivated by cooking. Sec-
ondly, however, and most importantly, if we assume that the
reported toxicity values for saponaceolide B and M in mice
hold also for humans, a 70 kg person would need to ingest the
overwhelming amount of 150 kg fresh mushrooms to reach
the LD50 value for saponaceolide B, and over 240 kg in the
case of saponaceolide M![15] Although such mushroom loads
are derived from simple body weight-based calculations, if the
stricter assumption is applied that body-surface area (BSA) scal-
ing might be better suited to translate toxicity values from
mice to humans,[16a] the required dose of mushrooms to be
consumed to reach the reported LD50 value for saponaceolide
B and M would result in 12.1 and 19.6 kg fresh T. terreum, re-
spectively, which is still far beyond any reasonable level of in-
gestion for any avid mushroom aficionado, even on repeated
sittings! Anyway, such dosages are to be considered with cau-
tion, since the BSA scaling approach for animal-to-human
dosage conversions is currently being debated among the sci-
entific community, and it has been recently suggested that
BSA-based calculations are not appropriate to derive interspe-
cies equivalents for dosages of natural products.[16b]
Therefore, it is most unlikely that saponaceolide B and M
might exert any toxicity to humans at the levels which are
found in T. terreum. Further experimentation is required to rule
out the possibility, if any, of long-term effects of saponaceolide
B and M, for example, after prolonged administration, provided
that their thermal stability to cooking is ascertained first.
In our opinion, therefore, the status of poisonous mushroom
that Liu and his team have attributed to T. terreum in their
paper is most questionable and, based on the above consider-
ations, their speculations on the alleged toxicity of T. terreum in
humans are readily disproved. Likewise, we find it unaccepta-
ble to point the finger of blame at well-known T. terreum as
the culprit for cases of mushroom-induced rhabdomyolysis
that have been described after consumption of the morpho-
logically well distinguishable T. equestre.
Regrettably, however, by virtue of its misleading title the
paper by Yin et al. about T. terreum has been already taken up
with eye-catching headlines by popular online science maga-
zines[17] and awareness has been also raised among field my-
cologists,[18] so that such a piece of information is unjustifiedly
alarming fungal collectors and mycological inspectors. We do
think that unsubstantiated information regarding edibility of
2  2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
ÝÝ These are not the final page numbers!

fungal species is most dangerous and we are concerned that it
might ultimately prompt regulatory bodies to ban T. terreum
from the positive list of edible and marketable species, thus
unduly hampering trade and usage of this mushroom species
in selected markets.
In conclusion, we feel the urge to reassure the mycological
community that T. terreum, on the basis of the current body of
knowledge, is not a poisonous species. At the same time we
are convinced that, in order to prevent new cases of mush-
room-induced rhabdomyolysis, mycologists and the scientific
community should always recommend to avoid consumption
of high amounts of mushrooms in consecutive meals for all
edible fungal species.[19]
Keywords: edible mushrooms · natural products
rhabdomyolysis · toxicology · Tricholoma equestre
[1] Taxonomically, T. terreum is placed by most modern authors within sec-
tion Terrea,[2] a group of species with predominantly grey cap colours,
and it is currently considered in a broad sense, that is, including similar
taxa that were often treated as distinct species in the recent past (e.g.
Tricholoma myomyces (Pers.) J.E. Lange, T. gausapatum (Fr.) Qul.); see
M. Christensen, J. Heilmann-Clausen, The genus Tricholoma - Fungi of
Northern Europe, Vol. 4, Svampetryk, Tilst, 2013.
[2] a) M. E. Noordeloos, M. Christensen in: Flora Agaricina Neerlandica, Vol.
4 (Eds.: C. Bas, Th. W. Kuyper, M. E. Noordeloos, E. C. Vellinga), A. A. Bal-
kema, Rotterdam. 1999, pp. 107 – 148; b) M. Bon, Les Tricholomes et re-
ssemblants. Flore Mycologique d’Europe 2, Doc. Mycol. Mm. H. S. 2,
CRDP, Amiens, 1991.
[3] For instance, see: a) R. Courtecuisse, B. Duhem, Champignons de France
et d’Europe, Delachaux et Niestl, Paris, 2013; b) G. Eyssartier, P. Roux, Le
guide des champignons. France et Europe, ditions Belin, Paris, 2011;
c) B. Cetto, I funghi dal vero, Vol. 1, 15th ed., Saturnia, Trento, 2008; d) R.
Phillips, Mushrooms, Macmillan, London, 2006; e) M. Bon, Champignons
de France et d’Europe occidentale, Flammarion, Paris, 2004; f) E. Ger-
hardt, Der große BLV Pilzfhrer fr unterwegs, BLV Verlagsgesellschaft
mbH, Munich, 2001; g) R. M. Dhncke, 1200 Pilze in Farbfotos, AT Verlag,
Aarau, 1993.
[4] E. Boa, Wild edible fungi. A global overview of their use and importance
to people, Non-Wood Forest Products 17, FAO, Rome, 2004. Also avail-
able at www.fao.org/docrep/007/y5489e/y5489e00.htm (accessed on 27
December 2014).
[5] Relevant legislation is provided in the Supporting Information.
[6] a) A. E. Bessette, A. R. Bessette, W. C. Roddy, S. A. Trudell, Tricholomas of
North America: a mushroom field guide, University of Texas Press, Austin,
2013; b) R. Imazeki, Y. Otani, T. Hongo, Nihon no kinoko [Fungi of
Japan], Yama-key Publishers Co., Tokyo, 1988; c) X.-H. Wang, P.-G. Liu,
F.-Q. Yu, [Color atlas of wild commercial mushrooms in Yunnan], Yunnan
Science and Technology Press, Kunming, 2004; d) S. C. Teng, Fungi of
China, Mycotaxon, Ithaca (NY), 1996.
Chem. Eur. J. 2016, 22, 1 – 4 www.chemeurj.org
These are not the final page numbers! ÞÞ
Correspondence
[7] X. Yin, T. Feng, J.-H. Shang, Y.-L. Zhao, F. Wang, Z.-H. Li, Z.-J. Dong, X.-D.
Luo, J.-K. Liu, Chem. Eur. J. 2014, 20, 7001 – 7009.
[8] Saponaceolide B was originally described from fruiting bodies of Tricho-
loma saponaceum (Fr.) P. Kumm.; see: a) M. De Bernardi, L. Garlaschelli,
L. Toma, G. Vidari, P. Vita-Finzi, Tetrahedron 1991, 47, 7109 – 7116; b) K.
Yoshikawa, M. Kuroboshi, S. Arihara, N. Miura, N. Tujimura, K. Sakamoto,
Chem. Pharm. Bull. 2002, 50, 1603 – 1606.
[9] R. Bedry, I. Baudrimont, G. Deffieux, E. E. Creppy, J. P. Pomies, J. M. Rag-
naud, M. Dupon, D. Neau, C. Gabinski, S. De Witte, J. C. Chapalain, P.
Godeau, N. Engl. J. Med. 2001, 345, 798 – 802.
[10] a) Z. Chodorowski, W. Waldman, J. Sein Anand, Przegl. Lek. 2002, 59,
386 – 387; b) Z. Chodorowski, J. Sein Anand, M. Grass, Przegl. Lek. 2003,
60, 309 – 310; c) Z. Chodorowski, M. Sznitowska, M. Wiśniewski, J. Sein
Anand, W. Waldman, A. Ronikier, Przegl. Lek. 2004, 61, 351 – 352; d) Z.
Chodorowski, J. Sein Anand, M. Madaliński, B. Rutkowski, B. Cylkowska,
P. Rutkowski, M. Wiśniewski, A. Hajduk, Przegl. Lek. 2005, 62, 468 – 470;
e) F. Assisi, S. Balestreri, R. Galli, Funghi velenosi, Dalla Natura, Milano,
2008; f) J. Sein Anand, P. Chwaluk, M. Sut, Przegl. Lek. 2009, 66, 339 –
· 340.
[11] Colour pictures of Tricholoma terreum and T. equestre are available for
comparison in the Supporting Information.
[12] a) P. Nieminen, A.-M. Mustonen, M. Kirsi, Food Chem. Toxicol. 2005, 43,
133 – 138; b) P. Nieminen, M. Kirsi, A.-M. Mustonen, Exp. Biol. Med. 2006,
231, 221 – 228; c) P. Nieminen, V. Krj, A.-M. Mustonen, Food Chem. Toxi-
col. 2008, 46, 781 – 786; d) P. Nieminen, V. Krj, A.-M. Mustonen, Food
Chem. Toxicol. 2009, 47, 70 – 74.
[13] a) P. Chwaluk, Przegl. Lek. 2013, 70, 684 – 686; b) N. B. Akıllı, Z. D.
Dndar, R. Kçyl, Y. K. Gnaydın, B. Cander, J. Acad. Emerg. Med. 2014,
13, 212 – 213.
[14] a) N. Sitta, M. Floriani, Econ. Bot. 2008, 62, 307 – 322; b) N. Sitta, P. Davoli
in Edible Ectomycorrhizal Mushrooms (Eds.: A. Zambonelli, G. M. Bonito),
Soil Biology, Vol. 34, Springer, Berlin, 2012, pp. 355 – 380.
[15] For the sake of comparison, a-amanitin, a potent fungal toxin responsi-
ble for severe—and often fatal—mushroom intoxications worldwide,
displays an estimated LD50 value of 0.1 mg kg 1 in humans and is found
at concentrations of 0.75–2.3 g kg 1 dry weight in fungal tissues of poi-
sonous Amanita species such as A. phalloides (Vaill. ex Fr.) Link and A.
bisporigera G.F. Atk., thus making the ingestion of a single fruiting body
sufficient for a potentially lethal intoxication; for selected references,
see the Supporting Information.
[16] a) S. Reagan-Shaw, M. Nihal, N. Ahmad, FASEB J. 2008, 22, 659 – 661;
b) O. L. Blanchard, J. M. Smoliga, FASEB J. 2015, 29, 1629 – 1634.
[17] H. Jia, Fatal toxins found in ‘edible’ wild mushrooms. Available at
http://www.rsc.org/chemistryworld/2014/06/fatal-toxins-identified-
edible-tricholoma-terreum-equestre-wild-mushrooms; http://
www.scientificamerican.com/article/fatal-toxins-found-in-edible-wild-
mushrooms/ (accessed on 27 December 2014).
[18] G. Kibby, Field Mycology 2014, 15, 110.
[19] L. Tofani, Difesa Sociale 2003, 82, 47 – 58.
Received: December 28, 2014
Revised: January 9, 2016
Published online on && &&, 0000
3  2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim & &
 Correspondence

CORRESPONDENCE
& Natural Products To eat or not to eat: Recent findings
casting doubts over the edibility of the
P. Davoli,* M. Floriani, F. Assisi, K. Kob, European mushroom Tricholoma ter-
N. Sitta reum are questioned on the basis of
&& – && mycological and mycotoxicological con-
siderations. Accordingly, T. terreum
Comment on “Chemical and should remain listed among edible
Toxicological Investigations of mushroom species.
a Previously Unknown Poisonous
European Mushroom Tricholoma
terreum”

& & Chem. Eur. J. 2016, 22, 1 – 4 www.chemeurj.org 4  2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

ÝÝ These are not the final page numbers!