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Jochen Gartz Et Al (1994) Ethnomycology Biochemistry & Cultivation of Psilocybe Samuiensis - A New Psychoactive Fungus From Koh Samui Thailand

Jochen Gartz Et Al (1994) Ethnomycology Biochemistry & Cultivation of Psilocybe Samuiensis - A New Psychoactive Fungus From Koh Samui Thailand

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ELSEVIER
Journal of Ethnopharmacology 43 (1994) 73-80
Ethnomycology, biochemistry, and cultivation of
Psilocybesamuiensis
Guzmk, Bandala and Allen, a new psychoactivefungus from Koh Samui, Thailand
Jochen Gartza, John W. Allenb, Mark D. Merlinc*
aKAI e. V./WIP. Department of Fungal Biorransjormation. Permoserstrasse IS. D-04318 Leipzig, GermanybP.O. BOX 12053. Honolulu. HI 96828-1053, USADepartment of General Science, University of Hawai’i. Dean Hall 2, 2450 Campus Road, Honolulu, HI 96822, .USA
Received 30 September 1993; revision received 19 January 1994; accepted 24 March 1994
Abstract
Several specimens of
Psifocybe
and
Cop&ndiu
species in Koh Samui, Thailand were recently collected for herbariumdeposit and scientific study. This paper presents an ethnomycological and biochemical study of one of the species;
P.samuiensis
GuzmBn, Bandala and Allen, a new psychoactive gill fungus reported from Thailand. Mycelium for thecultivation of
P. sumuiensis
was obtained on 6% malt agar from the spores of a dried specimen. The growth of
P.sumuiensis
was similar to that of
P. tumpunensis
Guzmin and Pollock, but more rapid than the mycelium of
P.semifanceafa
(Fr.: Sacc.) Kumm. Laboratory analyses indicates that the alkaloid content in cultured fruit bodies of
P. samuiensis
is of the same order of magnitude as that found in naturally occurring mushrooms of this species. HPLCanalyses of both naturally occurring and in vitro cultivated fruit bodies of
P. samuiensis
revealed high concentrationsof psilocybin and psilocin. Small amounts of baeocystin were also detected. Psilocybin levels varied from 0.23% upto 0.90%. The psilocybin content was highest in the caps. Psilocybin was also found in the cultured non-bluing myceliaof
P. samuiensis
and varied from 0.24% to 0.32% dry weight. The relative alkaloidal content of psilocybin, psilocin,and baeocystin found in
P. sumuiensis
was similar to that measured in many other psychoactive fungi species, but com-pletely different from that found in
P. semiianceuta.Keywords: Psilocybe sumuiensis;
Psilocybin; Psilocin; Cultivation; Koh Samui;
Psilocybe
spp.; Psychoactive fungi
1. Introduction
Recent ethnomycological investigations on Koh
Samui Island in Thailand (Allen, 1991;
Allen andMerlin, 1992a, b; Guzmin et al., 1993) confirm
l
Corresponding author.
reports that certain species of psychoactive fungiare ingested for recreational purposes by foreigntourists and some indigenous people
(Allen et al.,
1992).Koh Samui (280 km2 located at latitude 10” N,longtitude 100” E), is a small tropical island in theGulf of Siam where psychoactive fungi are
0378-8741/94/$07.000 1994 Elsevier Science Ireland Ltd.
All rights reservedSSDI 0378-8741(94)01135-M
 
14
J. Cart: er ul. /J. E~lmopharmacol. 43 (1994) 73-80
harvested by local farmers and their children. Thepsychoactive dung fungi
(Psilocybe cubensis
(Earle) Singer and/or
Psilocybe subcubensis Guz-
man) are known locally as ‘hed keequai’ (literally,‘mushrooms which appear after water buffalodefecates’). These mushrooms are sold by somefarmers or their families directly to tourists andresort restaurants. At some resort restaurants thefungi are offered in a variety of meals (Allen andMerlin, 1992a).Fungi specimens collected on Koh Samui Island(1991) for herbarium deposit and scientific studyinclude the following species:
P. cubensis, P. sub-cubensis, Copelandia cyanescens
(Berkeley etBroome) Singer, and a previously unreported blu-ing
Psilocybe
species,
P. samuiensis
Guzman, Ban-dala and Allen, sp. nov. (Fig. 1).Chemical investigations of the psychoactive pro-perties of some of these fungi include an un-published study by Stijve (Nestec Laboratories,Vevey, Switzerland) who analysed Koh Samui col-
Fig.
I.
Fresh harvested specimens of
fsilocybe samuiensis Guz-
m8n. Bandala and Allen. Ban Hua Thanon. Koh Samui.Photograph by John W. Allen.
lections of
P. cubensis
and
P. samuiensis,
as well asa Swiss collection of
P. semilanceata
(Fr.: Sacc.)Kumm. Tryptamine indoles were found in all threespecies. A second study by the authors was laterconducted utilizing naturally occurring materialand material grown only in vitro. Results of theHPLC and TLC analysis of both studies of
P.samuiensis
revealed high concentrations of tryp-tamine indole alkaloids. Furthermore, we weresuccessful in isolating a pure strain of
P. samuien-sis
on malt agar and grass seed.2.
Description of the cap, gills and stipe of
P.smuiensis
Guzmain, Bandala and Alien, sp., nov.2.1.
Cap
The cap is 7- 15 mm in diameter, subconvex toconic-convex, conic umbonate or campanulate-umbonate, frequently with a small papilla. It isviscid with a separable pellicle, even and striate tosulcate at the margin, and hygrophanous. It ischestnut or reddish-brown to straw-color, becom-ing pale straw-color or brownish clay when dry.2.2.
Gills
The gills are adnate to adnexed, clay color,becoming violaceous brown or chocolate brown-violet when dry, with white edges.2.3.
Sripe
The stipe is 40-65
x
1.52 mm, equal or slightlysubbulbous. It is hollow, white or whitish to palestraw color and covered with white fibrils. It iscontext concolorous with pileus, bluing withslightly farinaceous taste and odour.2.4.
HabitatP. samuiensis
was first collected in soil of mixedsand and clay, among fan palms in rice paddiessituated 2 km west of the village of Ban HuaThanon, Koh Samui, Thailand. Unlike
Copelan-dia.
and some species of
Psilocybe
which arecoprophilous,
P. samuiensis
does not fruit directlyin manure but appears scattered or gregarious inthe manured soil of rice paddies. This fungus fruitsfrom early July through late August.
 
J. Garr-_ CI al. /J. Ethnopharmacol. 43 (1994) 73-8075
3. Methodology
While foraging for psychoactive fungi on KohSamui island in August of 1991, John W. Allen,accompanied by several Samui children, harvested
P. samuiensis
for herbarium deposit and scientificexamination (see Guzman et al., 1993).
P. samuien-sis
first attracted the attention of the collectorbecause of its macroscopic similarity to
P.semilanceata
(the ‘liberty cap’ mushroom). Bothspecies can be found in similar environments (i.e.,pasture lands, rice paddies, etc.), occurring in themanured soil of ruminants. Carpophores of
P.samuiensis, P. cubensis
(and/or
P. subcubensis
which is macroscopically indistinguishable from
P.subcubensis),
and C.
cyanescens
were photograph-ed in situ, all growing in a single rice paddy.Psychoactive fungi species described in thisstudy were harvested from manured soil or decom-posed manure of Asian water buffalo
(Bubalusbubalis),
and cattle
(Bos indicus
and
Bos sun-duicus).
All of the above mentioned fungi were col-lected (2-8 August 1991) in rice paddies at fourdifferent locations near the villages of Ban Saket,Ban Hua Thanon, Bo Phut and Ban Lipa Yai onKoh Samui Island.Several collections of these fungi (including
P.samuiensis,
labeled as collection F), were sun driedand forwarded to Dr Gaston Guzman of the In-stituto de Ecologia, in Xalapa, Mexico forbotanical identification and to Dr T. Stijve ofNestec Ltd., Vevey, Switzerland for chemicalanalyses to determine the presence or absence oftoxic and/or psychoactive alkaloids.4.
Experimental
Mycelium was obtained from the spores of adried specimen of
P. samuiensis
by methodsdescribed by Stamets and Chilton (1983). It wasthen stored as stock culture on 6%) malt agar.Strains of a related species
P. tampanensis
Guzminand Pollock and
P. semilanceata
from Germanywere also obtained on agar. In a ratio of l-6% onmalt agar, the whitish mycelium of
P. samuiensis
grew at a faster pace than that of similar myceliumof
P. semilanceata.
The rapid growth of
P. tam-panensis
was similar to the growth of
P.
samuien-
sis; however, the former species soon formedbrownish sclerotia on the agar. Stamets andChilton
(I
983) reported similar growth patterns ofsclerotia on the agar of a related species
P. mex-icana
Heim. Even after a relatively long growthperiod (3 months), the mycelium of
P. samuiensis
formed only a few small brownish sclerotia (in theagar only, not in the culture).Similar growth patterns were also observedwhile cultivating the three species on
Lolium
seed/water (1:1.5), and in complete darkness.Observations on the rapid formations of sclerotiain
P. tampanensis
after a few weeks of cultivationwas first reported by Stamets and Chilton (1983).In contrast,
P. sumuiensis
under cultivation onlyformed thick whitish mycelium throughout themedia (rhizomorphs, diameter 2-3 mm), and pro-duced no sclerotia. Under the same conditions ofcultivation,
P. semilanceata
grew slowly, produc-ing only a fine and whitish mycelium with no for-mation of sclerotia or rhizomorphs.Psilocybin was found to be present in thecultured, non-bluing mycelium of
P. sumuiensis
grown on 6% malt agar. Amounts of psilocybin,ranging from 0.24O/o o 0.32% dry weight, wereanalysed in 5 different batches of mycelium grownover a 4-week period. Analyses also revealed thatthese quantities of psilocybin were much lowerthan those detected in the naturally occurring fruitbodies obtained from the field. Interestingly, noother indole derivatives were detected in theextracts of the in vitro grown mycelium.The alkaloidal levels obtained from the slightlybluing sclerotia of
P. tampanensis
were relativelyhigh when compared with the sclerotia of otherpsychoactive fungi species. In addition, theamount of psilocybin obtained from five differentcultivated samples of
P. tampanensis
grown on 6%malt agar and
Lolium
seed ranged from 0.34% to0.68% by dry weight, and from 0.41% to 0.61% inthree sections of sclerotia obtained from a singlecultivation on
Lolium
seed. The sclerotia of
P.tampanenesis
obtained from malt agar contained0.21-0.52% psilocin, but no baeocystin wasdetected. The sclerotia obtained from
Lolium
hada concentration of psilocin of 0.1 l-0.32%. Untilnow, no cultivation of complete fruit bodies of
P.sumuiensis
on either malt agar or
Lolium
seed has

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