The publ i s he r does not advocate the br eaki ng of the
law. The material herein is present ed as i nformat i on
which should be avai l abl e to the public. DEDICATION This book is respect ful l y dedicated to R. Gor- don Wasson and Albert Hofmann, whose inves- tigations of the botany and chemistry of the magic mushroom brought psilocybin to the world. "At last you know what the ineffable is, and what ecstasy means." -R.G. Wasson, 1972 Copyright 1976 by And/Or Press P.O. Box 2246 Berkeley, CA 94702 ISBN: 0-915904-13-6 First Pri nt i ng Layout : C. Schnabel TABLE OF CONTENTS FORHWORD 7 INTRODUCTION 11 STEP I: Locat i ng and Ide nt i f yi ng the Fungus: Col l ect i ng and Ger mi nat i ng Spores 17 STEP II: Growing Stock Inocula 25 STEP III: Gr owi ng on Sterilized Rye 33 STEP IV: Casing 45 STEP V: Har ves t i ng, Pr eser vi ng, and Dosage 51 AF TERWORD 55 CONVERSION TABLE 56 CHRONOLOGY 57 BIBLIOGRAPHY 61 GLOSSARY 62 FOREWORD Less t han t went y years have passed si nce Al be r t Hof- mann isolated and named t he hallucinogen psi l ocybi n. Hof- niann's psilocybin was ext ract ed from various species of mush- rooms whose occur r ence and ritual use in t he mount ai ns of Oaxaca had been discovered by Gordon and Val ent i na Was- son in the summer of 1953. Of the many species which were in use in Oaxaca, subsequent laboratory tests revealed t hat only one species was easily grown and able to f r ui t under a variety of art i fi ci al conditions. That one species is Stropharia cubensis the st arborn magic mushroom. This book is a path to this mushroom; how to grow it and how to pl ace it in your life like the shining light that it is. The sections which fol l ow give precise no-fail i nst ruct i ons for growing and preservi ng the magic mushroom. We have made these i nst r uct i ons as clear and direct as possible; what is described is only slightly more complicated t han canning or maki ng jelly. These i nst ruct i ons can be adapt ed to undert aki ngs of any size from a few jars to thousands. But bef or e all these details t her e should come a chat about just what this is really all about. We imagine that if you are avidly reading t hi s book it is probabl y because you have t aken dried mushrooms or been exposed to fresh ones in Lat - in America, so we do not begi n wi t h readers unf ami l i ar wi t h the joys of mushroom tripping. Our i nst ruct i ons arc a combi- nat i on of research i nt o ot her people's met hods of cul t i vat i on and procedures which we developed, tested, and f ound usef ul ourselves. Not hi ng we recommend is unt ri ed by us. There may be other ways to carry on small-scale cul t i vat i on indoors but cither they are variations on our method that are less direct or they are u n k n o wn to us. Cul t i vat i on of Stropharia outside on compost is possible in the U.S. if t he l ocal t emper at ur e is warm t hrough t he growi ng season. But compost cul t i vat i on i s an art i n i t sel f and demands more space, more e f f o r t , and more public exposure t han our indoor method. Get t i ng in- volved in composting a t on of manur e is not a necessary part of producing huge quant i t i es of per f ect magic mushrooms! Our method is scientific but our opi ni ons about Stro- pharia cubensis are not . Our opinions in t hi s mat t er do not rest upon the opinions of ot hers nor upon anyt hi ng wr i t t en 7 i i i a n y book, i nst ead t hey rest upon the exper i ence of the mushroom ps i l oc ybi n at t he 10 mg level; at t hat level a pecu- l i a r phenomenon occurs. It is the emergence of an I-Thou re- l at i onshi p bet ween t he person t aki ng t he psilocybin and t he ment al st at e it evokes. J ung calls this "transference 1 " and it was a necessary condi t i on of early and primitive huma ni t y' s r el at i ons hi p to its gods and demons. The mushroom speaks, and our opi ni ons rest upon what i t t el l s el oquent l y of i t s el f in t he cool night of t he mi n d : '"I am ol d, ol der t han thought in your species, whi ch is i t sel f fi ft y t i mes ol der than your hi st or y. Though 1 have been on earth for ages I am from the stars. My home is no one pl anet , for many worlds scat t ered through the s hi ni ng disc of the gal axy have conditions whi ch al l ow my spores an oppor- t uni t y for life. The mushr oom which you see is t he part of my body given to sex t hr i l l s and sun bat hi ng, my t r ue body is a f i n e network of fibers growing through t he soil. These net - works may cover acres and may have far more connections than the number in a huma n br a i n. My mycelial net wor k is nearly immortal-only the s udde n toxification of a planet or t he explosion of its parent star can wi pe me out . By means impossible to expl ai n because of cert ai n mi sconcept i ons in your model of r eal i t y al l my mycelial net works in the galaxy are in hyperl i ght communi cat i on across space and t i me. The mycelial body is as f r agi l e as a spider's web b u t t he collective hypermi nd and memory is a vast hi st ori cal archive of t he ca- reer of evolving i nt el l i gence on ma ny worl ds in our spi ral star swarm. Space, you see. is a vast ocean to those hardy l i fe forms that have the abi l i t y to reproduce from spores, for spores are covered wi t h t he hardest organi c s ubs t ance known. Across t he aeons of time and space dr i f t ma ny spore-formi ng life-forms i n suspended a n i ma t i o n f or mi l l i ons of years unt i l contact i s ma d e wi t h a s ui t abl e e n v i r o n me n t . Few such species are mi n d e d , onl y mysel f and my r ecent l y evol ved near rela- t i ves have achieved the hyper -communi cat i on mode and mem- ory capaci t y t ha t makes us l eadi ng members in t he communi t y of galactic i nt e l l i ge nc e . How t he hyper communi cat i on mode operates is a secret whi ch wi l l not be lightly given to man. But t he means should be obvi ous: it is the occurrence of psilocy- bin and psilocin in t he bi osynt het i c pat hways of my living body t hat opens for me and my symbi ot s t he vi si on screens to many worlds. You as an i ndi vi dual and man as a species are 8 on the br i nk of t he format i on of a symbiotic r el at i ons hi p with my genet i c material that will event ual l y carry humanity and earth i nt o the galactic mainstream of t he hi gher ci vi l i za- tions. "Since it is not easy for you to recogni ze ot her var i et i es of intelligence around you, your most advanced theories of politics and society have advanced only as far as the notion of collectivism. But beyond the cohesion of t he members of a species into a single social organism t her e lie richer and even more baroque evolutionary possibilities. Symbiosis is one of these. Symbiosis is a relation of mut ual dependence and posi- t i ve benefits for bot h of t he species i nvol ved. Symbiotic rela- t i onshi ps between myself and civilized forms of hi gher ani- mals have been established many times and in many places throughout the long ages of my devel opment . These relation- ships have been mut ual l y usef ul ; wi t hi n my memory is the knowledge of hyperlight drive ships and how to build t hem. I will t rade this knowl edge for a free t i cket to new worlds around suns younger and more stable than your own. To se- cure an eternal existence down the long river of cosmic time I again and again offer this agreement to higher beings and thereby have spread t hroughout the galaxy over the long mll- lenia. A mycelial net wor k has no organs to move the world. no hands; but higher ani mal s with mani pul at i ve abi l i t i es can become partners with t he star knowl edge within me and if they act in good f ai t h, ret urn both themselves and their humbl e mushroom t eacher to the mi l l i on worlds all ci t i zens of our starswarm are heir to." INTRODUCTION It seems characteristic of the human condition t hat man, in whatever environmental or existential mi l i eu he finds him- self, experiences an urge to seek cont act wi t h the essential myst ery underlying the fact of being. Indeed, the ent i re odys- sey of our species, both phyl ogenet i c and historical, can be seen as a groping toward some sensed transcendent fulfill- ment. The story of manof his art , science, philosophies, civ- ilizations and rel i gi onsi s largely the story of this quest for contact with the holy, numi nous, and self-transcending. It is a quest at least as old as man; evi dence indicating that early man possessed religious consciousness has been f ound dating back to the Mi ddl e Paleolithic. The archeological evidence shows clearly: Man was at home with the concept of the sa- cred long before he possessed writing, agr i cul t ur e, civilization. or science; it is a concept t hat has abided in the mind of man and guided him forward since the earliest i nfant hood of hu- manity, contemporary with, possibly even preceding, his ear- liest use of tools, fire, even language itself. The life of pre-literate man is one in which nat ure exists as the primary condition of existence: one is surrounded by it, one is immersed in it, one depends upon it for one's very survival. The quest for food and for the material necessities of life must be a constant and unending one for man-in-nature, a quest in which every pl ant and animal t hat one encounters comes under the scrutiny of a restless curiosity. Given this si t uat i on, it was inevitable that'sooner or later in the search for food man would accidentally ingest certain plants contain- ing compounds affect i ng the central nervous systemand find himself suddenly transported to a realm of the profoundest rapture and strangeness. Indeed, the ethno-mycologist R. Gor- don Wasson (1958, 1961) has suggested that the accidental ingestion of an hal l uci nogeni c plant, probably a mushroom, constituted man's earliest encount er wi t h the numi nosum, and led directly to the formation of the concept of deity and the super nat ur al . This not i on is not without a certain logical appeal: it st ands to reason that man's restless, roving eyes, scanning nature for potential sources of food, would quickly single out the lowly mushroom, so odd in appearance and so unl i ke t he rest of the veget at i on with which he was familiar. 11 Given a few t housand years for r andom experi ment at i on (a r el at i vel y short time in t he scale of prehistory), he would even- tually discover and ingest fungi containing centrally-active compounds, undergo t he hallucinogenic experi ence and the connection with the numinosum would be established. The scenario described is, of course, imaginary. We can- not know the exact circumstances under which man fi rst con- f r ont ed the psychedel i c experience. We do know, t hanks to t he work of Wasson and his colleagues in t he 1950s (cf. V.P. & R.G. Wasson, 1957, R.G. Wasson & R. Heim, 1958, & Was- son, 1957), t hat a religious cult centered around the ri t ual in- gcstion of hallucinogenic mushrooms has existed in the high- l ands of cent ral Mexico at least since before the Conquest, and is very likely much more ancient than t hat , its real origins hav- ing been lost in the mists of prehistoric time. But the fact re- mains t hat , whet her encountered through the ingestion of a fungus or some other pl ant , or through some spont aneousl y triggered altered st at e of consciousness, the direct experi ence of the transcendent has had and is having a profound impact on human history, per haps even on human evol ut i on. The urge toward the t r ans cendent and the dynami c tension that exists between the drive to t ranscend and the mundane neces- sities which impose themselves on t he primary f act of biologi- cal b e i n g i s in a sense what all history, all religion, art, philo- sophy, discovery and sciencein short, all of human thought and ci vi l i zat i oni s about . The urge to reach be yond t he known to what is unknown and unpl umbed is irredeemably woven into the fabric of human history. It is t hi s urge which bui l t the pyramids, Stonehenge and t he Gothic cathedrals. The same urge drove frail ships across the trackless oceans to t he shores of a new world, and t he same urge in our own t i me has driven us to fling a tiny bubbl e of light and air across the vast and howling abysses of space (t ha t cosmic mi l l i -mi cron) t hat separates our ear t h from its moon. It is t he same urge t hat stirs the shi ver along our spines when we gaze with wonder and longing at the star-dusted sky on a clear winter's evening. Today, we stand on the t hreshol d of the stars. Slowly it is emerging in mass consciousness t hat the next evol ut i onary step forward will so transform humani t y t hat all t hat has gone before will seem but a prelude. We stand at t he edge of history ready to accelerate our human exper i ence out i nt o t he vast chasm of night which engulfs our pl anet , the lessons of our 12 historical career still echoing down the corri dors of time. We are about to embark on the greatest adventure we have ever known, one that wi l l change our very not i on of what it is to be huma n; yet we should not forget that between ourselves as we ascend the r amp of the starsliip and our mushroom munchi ng ancestor gazing into his Pal eol i t hi c fire lie only sec- onds of cosmic time. This book is essentially a how-to manual for those who have the interest, t i me, and pat i ence required for cultivating "the magic mushroom" in their own homes. It is for people who feel that they still may be able to learn something by ex- periencing the primordial visions of t hei r ancest ors, and feel it strongly enough t hat they are willing to invest a l i t t l e t i me , money and effort in order to realize t ha t vision. By "magic mushroom" is meant those mushrooms which are members of the genus Psilocybe, and the closely related genera 5Yro/;/j< 7/vfl, Conocybe, Pa/uteolus, and Copelandia, Certain members of these genera contain the compounds psilocybin (4-phosphory- l oxy-N,N-di met hyl t rypt ami ne) and psilocin (4-hydroxy-N,N- dimethyltryptamine) as the active hallucinogenic agents (Fig. 1) . These compounds cont ai n the basic indole structure char- acteristic of most hallucinogens found in nat ure (cf. Schultes, 1973, p. 17f f . ), i ncl udi ng various amides of lysergic acid (of which LSD is a semi synt het i c represent at i ve), N,N-dimethyl- t r ypt ami ne, hanni ne and its analogues, and ibogaine. The most not abl e except i on to t hi s basic st r uct ur e is mescaline, which chemi cal l y is 3,4,5-t ri met hoxyphenyl et hyl ami ne. and hence is in t he same class as amphet ami ne (a-methylphenyl- et hyl ami ne). The cul t i vat i on information in this book pert ai ns only to one species of magic mushroom, Stropharia cubensis Earle. (The mycologist Rolf Singer has recent l y reclassified t h i s spe- cies i nt o t he genus Psilocybe. Hence in some r ef er ences it is referred to as Psilocybe cubensis Earle ex. Singer.,) It is prob- abl e t hat wi t h appropri at e adapt at i ons the met hods outlined 13 here could be applied successfully to the cultivation of other species. Our exper i ence has shown, however, t hat Stropharia cubensis is the easiest to cul t i vat e. At t empt s at cul t i vat i ng other species have so far met with problems in maki ng t he f ungus "fruit," or produce mushrooms, t hat will require fur- ther work to overcome. Our l i mi t i ng the di scussi on to one species, however, is not as unf or t una t e as it may seem since Stropharia cubensis is not onl y one of the strongest of the hallucinogenic mushrooms, but also one of the most wide- spread and readi l y obt ai nabl e. In nat ur e, its habitat is cow- dung, and it can be found in past ures dur i ng rainy, warm parts of the year in regions as diverse as the Southeastern U.S. and Cambodia, Australia and Colombia. Unlike other psilocybin- containing genera, which with few except i ons are fairly re- stricted endemics, the di st ri but i on of the Stropharia cuben- sis is world-wide (cf. Pollock, 1975). In fact , since its pre- ferred habi t at is cow-dung, its ci rcumt ropi cal distribution has doubtless been encouraged, if not caused, by the world cattle i ndust ry. Amusingly enough, the Stropharia could be said to exist as a "weed"" on high-technology cat tie-raising cul t ures. This i nt i mat e association with man via his domesti- cated cattle has probably existed for as long as humani t y has possessed pastoral technology. The procedures outlined in this book, if followed with care and persistence, will work for Stropharia cubensis. The procedures can be carried out by anyone in their own home, with just a minimum of equi pment and a few supplies and common chemicals that are no more than moderately di f f i c ul t to obtain. No special training in mycology or microbiology is necessary. What is necessary is to follow t he instructions close- ly and carefully. The procedure described herein consists essentially of four major steps. It begins with spores and describes step-by- step i nst r uct i ons for growing full-size mushrooms from spores wi t hi n six weeks. The first step involves locating the f ungus, collecting and germinating the spores, and isolating t he my- cel i um, or fungal threads, obtained from the spores. The next step involves cul t i vat i ng the mycelium on agar, a solid nutri- ent , in order to use it for inoculation. In the third step, my- celium grown on agar is then grown on a sterilized medi um of whole rye grains. In the fourt h and final step the rye-grown mycelium is cased, or covered wi t h soil, a process t hat induces the production of mushrooms. This book describes each of these steps in de t a i l , and can be put into pr act i ce by anyone abl e t o read and carefully f ol l ow t he i nst r uct i ons, provided that they can obtain spores or specimens of Stropharia cubeti- 14 15 16 STEP 1 : LOCATING AND IDENTIFYING THE FUNGUS COLLECTING AND GERMINATING SPORES In the New Worl d, Stropiwria cubensis can be found in appropri at e habitats t hr oughout t he Southern U.S., all through the coastal regions of Mexico, and t hr oughout coastal and equatorial regions of South America. In the U.S., it has been reported from Texas, Louisiana, Al abama, Mississippi, Ar kan- sas, F l ori da, Tennessee and Georgia. It s di s t r i but i on would probably be even great er were it not for the f act t hat i t s en- vi ronment al r equi r ement s l i mi t it to r egi ons of mi l d t emper - at ures and high humi di t y. Because of its specific habi t at and si ngul ar appearance, Stropharia cubensis is one of t h e easiest mushrooms to locate and i dent i f y. As al r eady ment i oned, i t can be found growing out of cow-pics in past ures during rai ny warm seasons. Other dung-growing mushrooms may also be f ound in the same pas- t ure, but these bear l i t t l e r es embl ance to Stropharia. The fol- lowing bot ani cal descr i pt i on of Stropharia cubensis is t aken from Mushrooms of North America by Orson K. Mi l l er , Jr. (also, sec Color Section fol l owi ng page 32): Cap pale yellowish, vi sci d; persi st ent r i ng: b l u e - s t a i n i n g st al k. Cap 1.5-8 cm br oa d, conic, bel l -shaped, convex in age. viscid, wi t h o u t hai r s , whi t i s h t o pale yellow, light brownish i n age. stains bl ui sh i n age. Flesh f i r m, wh i l e , brui ses b l u e . Gills a dna t e ( a t - t a c h e d ) t o adnexed (not ched), close, gr ey t o vi ol e t -gr e y i n age wi t h wh i l e edges. St al k 4- 15 cm l ong, 4-14 mm i hi ck. enl ar gi ng somewhat t owa r d t he base, dr y, wi t h o u t hai r s , whi l e s t a i n i n g bl ue whe n br ui s e d. Veil whi t e , l eavi ng a super i or me mbr a nous r i ng. Spores 10- 17/ u x 7- I Oj U e l l i p t i c a l t o oval i n si de-vi ew, t hi ck-wal l ed, wi t h a large por e at apex, pur pl e-br own spore p r i n t . Cys t i di a ( s t e r - ile cells) on gill edge club-shaped wi t h rounded heads. Mi l l er places this species in the genus Psilocybe, af t er Singer. The flesh of t hi s mushroom exhibits the pr oper t y of s t ai ni ng a bl ui s h col or when bruised or broken. This blue- st ai ni ng r eact i on is appar ent l y an enzymat i c oxi dat i on of some indole subst r at e (t r ypt ophane, 5-hydr oxyt r ypt ami ne, or psilo- c ybi n) and is a fai rl y r el i abl e i ndi c a t or of t h e presence of psi - , not onl y in Strophuria cubensis, b u t al so in other 17 closely related genera (members of the family Strophariaceae) (cf. Benedict, et al., 1967). Other mushrooms, such as mem- bers of the genus Russula, section Nigricantinae, and Boletus, exhibit a similar blueing. The blueing in these cases, however, is not due to the presence of indole substrates and these mush- rooms otherwise bear no resemblance whatever to Stropharia cubensis or related species (Singer, 1958, p. 247). The blueing of the Stropharia and Psilocybe species is also accompanied by a strongly positive reaction wi t h the chemical reagent metol (p-methyl aminophenol), a common compound used in photographic darkroom work. This reagent dissolves in 20 times its weight of water, and will turn a deep purple color within 1-30 minutes when a few drops are added to a section of crushed stem. The compound in solution is unstable and must be used immediately after it is mixed with water. (Singer, 1958, p. 247;Enos, 1970, p. 5). Once one has located a specimen or specimens of Stro- pharia cubensis, and been satisfied as to its identity in all par- ticulars, it is necessary to collect spores for cultivation. The test with metol is more or less a double-check and is not really essential, since most specimens will readily blue when the stipe (stem) is broken. Spores can be easily collected in the following manner: Take one or more fresh specimens with the caps fully open; using a sharp kni fe, cut off the stipe as close to the gills as possible (cf. Fig. 3) and place the cap gill- side down on a clean sheet of white paper, and leave for 24 hours. It doesn't hurt to cover the caps with a small bowl while t aki ng the spore print in order to retard dessication. When the caps are removed, a dark-purplish, radially symme- trical deposit of spores will remai n on the paper where the gills contacted it. The paper should then be folded and sealed in an envelope in order to prevent furt her contamination by air-borne spores of other species of lower fungi. A single spore-print contains tens of millions of spores, and is suffi- cient to make hundreds of spore germinations. The following variation on this method was suggested to us as a way of enhancing the sterility of the spore print: Take four standard flat microscope slides, swab with alcohol, and flame in an alcohol flame or but ane torch (Fig. 2). On a clean flat surface, such as a table-top swabbed with Lysol, lay the slides side by side and end to end, so that they are arranged as in Fig. 3. Place the fresh cap in the exact middle of the slides 18 so that approximately % of the cap covers each slide (Fig. 4). Cover and wait 24 hours. When the cap is removed, the end of each slide will be covered with spores, and the slides can then be sealed, together or separately, in plastic or paper. Once the spore-print has been collected, it is necessary to germinate some spores in order to begin the life-cycle that will eventually culminate in the production of more mush- rooms. Before we outline procedures for germinating the spores, a brief discussion of the stages in the life-cycle of these higher fungi follows; readers who do not care to read this somewhat technical portion may skip to page 21, paragrph 3. All gilled fungi are members of the class Basidiomycetes, i.e., they are characterized by the production of spores on club-shaped appendages called basidia. Spores borne on basi- dia are called basidiospores. Most of the conspicuous fungi that one encounters, such as mushrooms, puffballs, and bracket fungi are members of the subclass Homobasidiomy- cetes. Of the members of this subclass, the gilled mushrooms are placed in the order Agaricales. The life-cycle of a typical homobasidiomycete is illustrated in the frontispiece. The ba- sidiospores germinate to form a monokaryotic hypha. A hypha is a tubular fi l ament ; an aggregation of these hyphae collectively comprise a mass of thread-like filaments referred to as the mycelium. The mycelium comprises the main body, or thallus, of the fungus. The stalked, capped structure which we call the mushroom is actually only the "fruiting body" or the spore-producing reproductive structure, and constitutes only a small portion of the total mass of the fungus; the great bulk of the organism exists underground in the form of a net- work of mycelium, which occasionally "fruits," or produces mushrooms, under appropriate conditions. The basidiospores germinate to produce a monokaryotic mycelium, i.e., a mycelium having only one nucleus per cell. This mycelium grows out until it encounters another mono- karyotic mycelium, germinated from another spore, that is a compatible mating type. If the monokaryotic mycelium does not contact a compatible monokaryotic mycelium, it eventu- ally dies. In situations where two compatible monokaryotic mycelia do make contact, however, a process called somato- gamy, or a fusing of the somatic cells of the two mycelia, takes place, but fusion of the nuclei does not take place. The result of somatogarny is the establishment of a dikaryotic my- 19 cel i um, i . e. , a mycel i um possessing t wo nucl ei , one from each of the monokaryot i c mycel i a, in each of its cells (cf . frontis- piece). The dikaryotic mycelium stage is the most prolonged por t i on of the life-cycle and is also t he main assimilative stage of the fungus. The di karyot i c mycelium can propagat e vege- tatively i ndef i ni t el y wi t hout going t hrough a sexual (spore- producing) stage. Given appropriate conditions, however, the di kar yot i c mycel i um can be i nduced to "fruit"; the undi f f er - entiated mycelial thallus of the f ungus begins to weave itself together into an art i cul at ed, spore-bearing "frui t i ng body." in this case, into a mushroom. The mushroom continues to en- large and t hr us t above the ground, incorporating more and more mycel i um whi l e at the same time expandi ng by absorp- tion of water. At a certain stage in the growth of the mush- room, or basidiocarp, club-shaped structures called basidia form on t he under si de of the gills. At t hi s point, karyogamy, or fusion of the two nucl ei of the dikaryotic mycelium takes place wi t hi n the basidia (cf . f r ont i s pi ece). This is the only diploid, or 2n, stage in the life-cycle of the fungus, and is also the bri efest stage, for meiosis, or reduction di vi si on of a dip- loid ( 2n ) nuc l e us t o 4 haploid (n) nucl ei occurs immediately following karyogamy (cf. front i spi ece). The result of meiosis is the production of four haploid nuclei within the basidium (cf. f r ont i spi ece); these are t hen pushed out of the basidium and become sur r ounded by hard sheat hs to form the basi di o- spores (cf . front i spi ece). The result is t he basi di um bearing four basidiospores on its outer surface as in the frontispiece. These basi di ospores eventually det ach from t he basi di um to begin the l i fe-cycl e again. Fungi of t he fami l y St rophari aceae, which i ncl udes Stro- phaiia cubensis and most other psilocybin-contahiing genera, are genetically compl ex with respect t o t he ma t i n g compat i - b i l i t y of di f f er ent monokar yot i c mycel i a. These fungi are hetemrliallic and tetrapolar, t hat i s, t hei r sexual cycl e is de- pendent on the fusion of two compatible monokaryotic my- celia, and their sexual compat i bi l i t y is governed by two sets of factors: In t et r apol ar het er ot hal l i s m. t wo s et s of f a c t or s , t he A' s and B's, are i nvol ved. If a sexually reproduci ng thallus is to be estab- l i s hed, somat ogamy mus t oc c ur bet ween mycel i a di f f er i ng i n bot h sets of f act or s for exampl e AB x ab. The n u mb e r of mat i ng 20 classes is somewhat great er than in bi pol ar forms, since f our t ypes t ypi cal l y arise from spores of a single basi di ocarp. Obvi ousl y these mating types, numbe r i ng i n the hundr e ds i n bot h bi pol ar and t et r apol ar species, cannot be designated as sexes! (Scagel, et a!., 1967, p. 69.) Keepi ng this i nformat i on pert ai ni ng to the sexual char- acteristics of these fungi in mind, let us return to the prob- lem of spore germi nat i on; the rel evance of our digression into the mat t er of life-cycle and sexual compat i bi l i t y will be seen shortly. Once one has obtained a spore print from Stfophana cu- bensis, the monokaryotic mycel i um can be easily obtained by germinating t he spores on an appropriate solid nut r i ent me- di um, such as Potato Dextrose Agar or Malt Extract Agar. A more detailed discussion of various kinds of nutrient agars and how to prepare them will be given below in t he section on Growing Stock Inocula. For t he present , however, simply assume t hat one has several clean, sterile petri plates which have been filled with an appropriate solid nutrient medium (see Fig. 5). Take the clean paper or slide on which the spore print is deposited, and using a clean knife, i nocul at i ng loop, or similar implement which has been sterilized by flaming in an alcohol lamp (Fig. 6), simply scrape the spore print lightly with the i mpl ement (Fig. 7), then transfer the adhering spores (Fig. 8) to the medi um by touching the surface in one or more spots with the tip of the implement (Fig. 9). Care should be taken to do this as quickly as possible, keeping the cover off the petri plate for the shortest time necessary, in order to minimize the chances of cont ami nat i ng t he pl at e wi t h the air- borne spores of contaminants. A variation on this method can also be used: Instead of scraping the spores directly onto the plate, they can first be scraped i nt o about 10 ml of sterilized wat er . Shake this vigorously, t hen di l ut e to 100 ml by adding sterile wat er. Using a sterilized pipette or syringe, take up 2-3 ml of diluted spore solution, and point-inoculate the petri plate by pl aci ng a drop of the sol ut i on at t wo or three separ- ate points on the pl at e. Allow t he covered i nocul at ed pl at e to st and at room t emper at ur e for 3-5 days. During this time, t he spores will germinate and monokaryotic mycelium will grow radi al l y out war d from each p o i n t of i nocul at i on. The plate should be l ef t undi st ur bed unt i l the mycel i um from t wo dif- 21 f er ent spores or two di f f er ent points of i nocul at i on has grown together and made cont act . A few days after cont act has oc- curred, one can be reasonably sure t hat somat ogamy has tak- en place and t hat a di kar yot i c mycelium has been established. In cul t i vat i ng the fungus to the frui t i ng stage, one works pri mari l y wi t h a single strain of dikaryotic mycel i um. How- ever, because spores of several di f f er ent mating types are pro- duced by a single mushroom, a petri pl at e i nocul at ed with spores will have possibly several dozen di fferent strains of di- karyotic mycel i um growing on i t . It is therefore necessary to isolate one of the strains so that one can grow out stock inoc- ula from a single, uni form strain. This can be accomplished using a scalpel, dissecting needle or inoculating loop in which the loop has been bent to form a hook (Fig. 8). The imple- ment is fi rst sterilized in an alcohol f l ame; then the petri plate is opened slightly and a very small piece of mycelial tissue is snagged on the end of the bl ade, needle, or hook, and trans- ferred r api dl y and deftly to a second clean sterile pet r i pl at e containing an appropriate solid nut r i e nt medi um. Di karyot i c mycelium is isolated using exactly the same techniques as are used in t ransferri ng mycel i um from one pet ri pl at e to anot her; for figures illustrating this procedure, see Step 11, Figs. 13- 16. By selecting a very small piece of tissue in t hi s way. one can be reasonably sure t hat only one strain of di kar yot i c my- celium is being removed and isolated. The di karyot i c myceli- um thus isolated from the spore-germination pl at e will grow outward in all directions from t he point of i nocul at i on on the new pl at e and should cover most of t he surface of the me- dium within 8-12 days. One can t hen go ahead and make fur- ther transfers to new plates with a fair degree of cert ai nt y t hat one is working wi t h a single strain of di kar yot i c mycel- ium. It is pr obabl y advi sabl e to isolate several di f f er ent st rai ns of di karyot i c mycelium onto separat e plates by t aki ng tissue from di fferent sections of t he spore-germination plate. Di f f e r - ent strains isolated from a single spore-germination pl at e should be identified by labels and compared for vigor of growth and vigor of f r ui t i ng abi l i t y, so t hat , t hr ough observa- tion and trial-and-error, t he strain showing ma xi mum vigor in bot h respect s can event ual l y be i dent i f i ed and used exdusi ve- y t hereaft er. Isol at i ng the most vi aorous strain t akes t i me and 22 23 careful observation: however, t hi s need not i nt er f er e wi t h con- t i nui ng on to t he second and third steps of the process, since any dikaryotic mycelium that has been properly isolated from other strains should exhibit frui t i ng abi l i t y. Af t er several dif- ferent strains have been put through several fruiting stages it should be apparent which st rai n is most vigorous. If one has fresh mushroom specimens, it is possible to employ anot her method of i sol at i ng dikaryotic mycelium wi t hout utilizing spores. This is the method of subcutaneous isolation (cf. Enos, 1970). Remove the stipe f r om a fresh mushroom, and, using three or four needles, fasten the cap gills down to a piece of cork-board or card-board. Swab the cap surface with t i nct ure of iodine using a sterile cotton swab. Then, using a flame-sterilized scalpel, remove a small section of the outer cuticle of the cap. Then resterilize t he scal pel , and remove a small piece of subcut aneous flesh from the cap, and t ransfer it to a sterilized nut r i ent plate. Since the flesh of the mushroom has been woven together out of dikaryotic mycelium, it will grow out across the pl at e in the same man- ner as mycelium isolated from a spore-germination plate. This procedure eliminates the step of having to isolate di fferent dikaryotic strains, since mycel i um isolated in this manner con- sists of only one strain. On the other hand, this procedure lim- its one to working with only one strain. 24 STEP II: GROWING STOCK INOCULA Once one or more st r ai ns of di karyot i c mycelia have been successfully isolated, it is necessary to build up a stock of mycelial cul t ures grown on sterile agar medi a. The inocula from t hi s stock will be used to inoculate the mycel i um ont o sterilized rye or other grai n. Before proceedi ng to t hi s step, however, it is advisable to have a good suppl y of inocula grown out on sterile agar medi a, so t ha t one will have pl ent y of sterile inocula even if a few cul t ures should succumb to contamination. The i nf or mat i on in this sect i on t herefore de- scribes procedures for preparat i on, st eri l i zat i on, and i nocul a- t i on of solid nut r i ent medi a. Most laboratory work wi t h higher f ungi , yeast s, molds, bacteria and so on involves growing t he organi sm on a solid agar me di um t o whi ch appropri at e n u t r i e n t s have been added. Agar is a pect i n-l i ke subst ance extracted from cert ai n ki nds of sea-kelp, which, when dissolved in boiling water and allowed to cool, solidifies to a gel at i nous consi st ency. Agar is a stan- dard i t em in al l microbiological wor k, and is avai l abl e from almost any sci ent i f i c supply company. It is also stocked by many heal t h-food stores and Oriental food stores as a dietary s uppl ement . Potato Dextrose Agar (PDA) and Mal t Ext r act Agar (MEA) are standard solid nut r i ent medi a sui t abl e for cultivat- ing the mycel i a of most hi gher f ungi , i ncl udi ng Stropharia cubensis. Bot h t ypes are commonly avai l abl e in a premi xed form from most sci ent i f i c suppl y compani es. The premixed type need only be dissolved in boi l i ng di s t i l l e d wat er . Usual l y about 1 5-20 g of pre-mixed agar me d i u m per 1000ml of wat er is used. The appropriate proportions and mixing instructions are usually pr i nt ed on the cont ai ner of dr i ed agar pr epar at i on. With very l i t t l e t roubl e, one can also manuf act ur e one's own PDA or MEA; a reci pe and pr ocedur e for each t ype is gi ven on t he f ol l owi ng page. 25 P.D.A. 250 g. potatoes 15 g. agar 10 g. dextrose 1.5 g. nut r i t i onal yeast (or yeast e xt r a c t ) Shred t he impeded potatoes i nt o a col l ander and t hen rinse them for t hi r t y seconds with cold t ap water. Combi ne the rinsed pot at oes with one liter of wat er and gently boil for thirty mi nut es. Filter the resulting pot at o broth t hrough mus- l i n or cheese cloth and discard the potatoes. To the l i t er of pot at o broth add the agar, dextrose, and yeast whi ch you have previously weighed out and mixed t oget her in a baggie. While stirring gently add in the mixed powdered ingredients. Gent l y boil for ten minutes or unt i l the solution is clear. Take care not to allow the solution to boil over. Add enough wat er to r et ur n t he t ot al volume of the solution to one liter. Pour the solution whi l e still hot into petri plates, baby food jars, or slant cul t ure tubes (Fig. 11) . Use just enough to cover the bot- tom of a plate or baby-food j ar to a dept h of about % in.; if using tubes, fill about ^full. The sol ut i on may be allowed to cool or sterilized i mmedi at el y. Sterilization procedures will be described shortly. A recipe for Mal t Extract Agar (MEA) follows: To 1 liter of gently boiling wat er add a previously weighed and mixed powder containing: 20 g. malt or malt ext ract (may be powder or s yr up) 15 g. cornsteep liquor (opt i onal ) 20 g. agar 0.1 g. Potassium phosphate dibasic (K 2 HPO 4 ) 0.1 g. Calcium car bonat e (CaCO 3 ) (powdered oys- ter shell may be used) Corn-steep l i quor, whi ch is an optional i ngr edi ent , may be made by boiling dri ed corn in wat er, l et t i ng the broth stand for several days, then sterilizing it in a mason jar in a pressure cooker. Corn-steep l i quor made t hi s way shoul d be refriger- ated. The liquid mal t extract sold in the syrup sections of most grocery stores is qui t e suitable for this medium. Af t er the nut ri ent s have been completely dissolved in the wat er, the hot sol ut i on is poured into plates in the same manner as the PDA. It does not hurt to add 1.5 g of yeast ext ract or nu- tritional yeast to ei t her of the media described above. This provides an addi t i onal source of proteins and B vitamins. If some of the nut r i ent solution is l ef t over aft er pouri ng t he plates, t he flask may be sealed and stored in the refri gerat or i ndef i ni t el y, or sterilized with the plates and stored on the shelf. When one wishes to make more plates, the medium can be reliquified over heat and reused. The two t ypes of media described above are qui t e easy to prepare and will be suitable for growing stock inocula. It is a good idea to mix up and have on hand both types of medi a, and to use them alternately in prepari ng batches of plates. In this way the fungus will not become accustomed to one type of medium and thus will be forced to use di fferent parts of its genome in adapting to the different media. This will prevent the mycelium from succumbing to any "senescence factor" or tendency to age physiologically and t hus to lose vigor after a period of t i me. These t wo types of medi a are completely adequat e for growing out one's stock i nocul a. From a purel y practical st andpoi nt , we have found PDA and MEA to be easily and readily prepared from a relatively few common ingredients. Unless one wishes to get involved in compl ex nut r i t i onal stu- dies, it is unnecessary to bother wi t h other reci pes. Ot her types of media may be used, however, and those who do wish to get more deeply into this step of the process are urged to consul t Enos, 1970 (see Bibliography). Once one has prepared an agar medi um and poured it in- to the petri plates, baby food jars, slant cul t ur e t ubes, or ot her s ui t abl e recept acl es, it is necessary to sterilize the medi um in the receptacles in order to kill the spores of bacteria, yeasts, and ot her mokl s which get into t he me di um from the air. This can be done via t he f ol l owi ngpr ocedur e: If a l aborat ory aut o- clave is not avai l abl e, a st andard home cooki ng or canning pressure cooker can be used. We use and recommend the All American 94114 pressure cooker, avai l abl e from Whole Earth Catalogue (see Fig. 27). Pl ace a small amount of water ( a ppr ox. 1 l i t er ) in t he bot t om of t he cooker (tap water will 26 do) so that the surface is covered. Place t he recept acl es con- taining the medi um i nt o t he pressure cooker. Be sure to stack t hem car ef ul l y (see Fig. 12); a small enameled t r ay is useful for this. Not e: If using pr es t er i l i zed pl ast i c pl at es, pour the me di um i nt o the plates after st eri l i zi ng; do not autoclave plas- tic plates. It does not mat t er whet her t he medi um is still hot and liquid, or whet her it has been allowed to cool and sol i di f y, since t he heat of the s t er i l i zat i on process will r el i qui f y the medi um anyway. If baby food jars or cul t ure t ubes are used, be certain that the lids are left loose, not screwed down tight. when they are being sterilized. Seal the lid of t he pressure cooker, but leave the stopcock open. Bring the cooker to a boi l over high heat on a stove. When the water has begun to boi l vi gor ousl y, a good head of steam wi l l begin to vent through the st opcock; it should be closed at t hi s p o i n t , and the pressure allowed to build up to bet ween 15-20 Ibs. Then reduce t he heat just enough to mai nt ai n pressure at this level for 45 mi nut es to 1 hour . The st andard st er i l i zat i on time for solid medi a at these t emper at ur es (250 F . ) and pressures (15-20 Ibs. ) is 15 mi nut es, but experi ence has shown t ha t t hi s is of t en insufficient to insure complete sterilization. Allowing a strong head of steam to build up in t he pressure cooker be- fore closing t he st opcock is also important, for if it is closed prematurely, the pressure will rise but the water will be un- able to vapor i ze, and dry heat requires much longer to accom- plish st er i l i zat i on. Af t er the medi um has been st eri l i zed at t he correct pres- sure for 45-60 minutes, turn off the heat or car ef ul l y remove the cooker (r emember that the medi um is l i qui d at t hi s poi nt and can slosh a r ound) and allow the cooker to cool to room temperature before opening the stopcock; ot her wi se, t he sud- den release of pressure will cause the medi um to boil over. Whe n the cooker has cooled to room t emper at ur e or slightly above, open the stopcock and allow any excess steam to escape. Remove the lid and carefully remove t he recep- tacles cont ai ni ng the medi um. Place t he receptacles inside a pre-sterilized inoculating hood (see Fig. 10) or on a cl ean, smooth table-top which has been wiped down wi t h Lysol or similar strong di si nf ect ant . As the receptacles cool f ur t her , t he 28 fig. 10: Two vi ews of a homemade i n o c u l a t i n g hood. medi um will solidify. If sl ant cul t ur e t ubes are being used, they should be placed on an angle while the medi um is still liquid, to provide maximum surface area for mycelial growth. When t he receptacles have cooled compl et el y to room t emperat ure, and t he medium is ful l y sol i di f i ed, they are ready to be i nocul at ed. If possible, i nocul at i on should be ear- ned out inside an inoculation hood such as t hat shown in Fig. I. Commerical hoods are avai l abl e, or a home-made hood can be const ruct ed out of wood and al l j o i n t s sealed with silicon caul ki ng compound. Presterilize the hood before in- t r oduci ng the c ul t ur e r ecept acl es by spraying all i nsi de sur- ges t horoughl y with Lysol aerosol, or a mi xt ur e of 25% Chlorox-distilled wat er sol ut i on, or bot h. If a hood is not available, i nocul at i on can be carried out in t he open ai r in a room in which t he air is relatively s t i l l , i . e. , a room wi t hout any drafts. The air of the room should be sprayed beforehand 25% Clorox solution and the surface on which i nocul a- on s to be done wiped down wi t h a strong Lysol sol ut i on, tenle procedures should always be done weari ng l at ex soves which have been sterilized by spraying wi t h Lysol 29 Inoculation should be done using a disposable scalpel or an inoculating loop which has been opened to form a hook (Fig. 8), and can be carried out in essentially the same manner as was described for isolating dikaryotic mycelium from a spore ger mi nat i on pl at e. Select a completely st eri l e, vigor- ously growing culture from one's stock of di kar yot i c mycel- ium isolated from spores. The mycel i um of a vigorous cul- ture should be pure white and fluffy in appearance, and preferably less t han 10 days old (cf. Fig. 13). Wash one's hands and arms thoroughly in soap and wat er before begin- ning work, t hen wipe down wi t h alcohol. Wear thin latex gloves sprayed with Lysol or Clorox-water solution as added protection (Figs. 13 & 14); t al cum powder can be sprinkled inside first to make them easy to slip on. Wear a short-sleeved or sleeveless shirt for the process to avoid i nt r oduci ng con- t ami nant s from one's clothes. Pass the wire of the hooked in- oculating loop through the flame of an alcohol lamp until it has been heated to redness in all of its parts, but especially at the end (Fig. 15). Open the cul t ure just enough to insert the end of the loop, and snag a small piece of mycelial tissue or small plug of agar and t ransfer this rapidly to the freshl y steri- lized me di um, again opening the lid just enough to insert the inoculum (Fig. 16). Wi t hdr aw the loop and replace t he lid on the newly i nocul at ed culture. If a disposable scalpel is used instead of a loop, inoculation can be accomplished by cutting a small square (c. 1 mm) of mycelia-grown agar from the cul- ture pl at e, and transferring this to the new plate. If using tubes or baby food j ar s, the lids should be l eft fairly loose to allow for aerat i on. Repeat this process for as ma ny times as one has receptacles to inoculate. It is not necessary to use a new cul t ure for each i nocul at i on; a single culture is sufficient to inoculate dozens of fresh plates. After a batch has been in- ocul at ed, however, the cul t ur e used as t he source of i nocul a should be discarded. Let t he freshl y i nocul at ed medi um stand at room t em- perature for 3-5 days. Dur i ng this time t he pure whi t e, thread- like mycel i um will spread radially across the surface of the medi um, covering it compl et el y wi t h i n 7-20 days. Growth of the i nocul um shoul d be apparent by t he f our t h day aft er i n- oculation. Also apparent by about this t i me will be any con- 30 fig. 13: Materials for pl a t e -t o-pl a t e fig. 14: Di si nfect i ng r u b b e r gloves i nocul at i on. with Lysol. 31 t ami nant s that have got t en into the cultures during inocula- tion in spite of precautions. They usually appear as small white dots with blue-green centers, and grow much more rap- idly than the mycelium. These are usually other molds, such as Penicillium and Aspergillus, Neurospora or vari ous yeasts. Most are easy to distinguish from the mushroom mycelium, since the mycelium is pure white, occasionally with a slight tinge of bl ue, while the contaminants may be green, blue- green, black, yellow, dirty-gray, and so on, and otherwise do not resemble the mycelium. Any contaminated cultures should be discarded as soon as one is certain that contamina- tion is present. It is normal to lose a few cultures to contami- nat i on, and one should not be discouraged by it. It is practi- cally impossible, under non-laboratory conditions, to elimi- nate all contamination; but as one gains practice in making inoculations, speed and technique should gradually improve so that contamination can be held to a minimum. It would be a good idea to consult an introductory microbiology text for i nf or mat i on relating to fast and efficient inoculating tech- niques. 32 STEP III: GROWING ON STERILIZED RYE When a number of mycelial cul t ures have been success- fully grown on solid agar medi um, one is faced with a choice: It is possible to stop at this poi nt , and concentrate on perfect- ing techniques for mycelial growth on agar. The mycelium it- self cont ai ns psilocybin and can be ingested for hal l uci nogeni c effects. The amount of psilocybin present in t he mycel i um is det er mi ned by the richness of t he medi um on whi ch it is grown. Thus for an i ndi vi dual wishing only to obt ai n psilocy- bin from mycel i um, there is a wide-open area for investiga- tion; viz. , to discover a suitable nut r i ent medi um t hat gives a maximum yield of psilocybin per unit surface area of agar. If l aborat ory f aci l i t i es are avai l abl e, psilocybin can also be ob- tained from mycelium grown in liquid medium in shaken or submerged flask cul t ur es. Since the necessary equi pment for this is unavai l abl e to most peopl e, this approach is not dis- cussed f ur t her her e. Int er es t ed readers are urged to consult Cat al fomo & Tyl er, 1964. The other choice at this stage involves moving on to the third step in the procedure, whereby mushrooms can be ob- tained by inducing the mycelium to frui t . In order for vigor- ous fruiting to take place, the mycelium must fi rst be grown out ont o sterilized rye. wheat , barley or other similar grain, so t hat a mass of mycel i um weighing from 50-100 grams is obt ai ned. The growing of mushroom mycelium on st eri l i zed grain is a standard procedure in commercial mushroom cul- t ure t hat is used to produce "spawn" for inoculation i nt o beds of horse-manure compost. The procedures described in this section are i n f act a da pt e d, wi t h appropri at e modi fi cat i ons, from a process originally developed by San Ant oni o ( 1971) for growi ng f r ui t s of t he common edible mushroom Agaricus bisporus unde r laboratory condi t i ons. The steps involved in growing the mycelium ont o rye-grain medium are described below. While the mycelium will grow and fruit suitably on many t ypes of gr a i n, i ncl udi ng rye, wheat , bar l ey, triticale, Oa ts, brown rice, sorghum, mi l l et and even buc kwhe a t , our 33 experience is t hat rye works as good as any and is less expen- sive than most . Therefore we have pri mari l y worked with rye in this stage. One must be cer t ai n, however, that the rye used is packaged for human consumpt i on, and not grown as f eed; feed rye has usually been t reat ed with a fungicide. At t hi s stage, it is useful to construct a st yr of oam box with a window in the lid such as illustrated in Figs. 17-20. These boxes can be obtained from pet stores and tropical fish dealers and can be used as a convenient modular system for incubating jars in a high-humidity, constant t emperat ure environment. To prepare the rye medium, begin with a clean, wi de-mout h quart mason jar with a dome and ring lid. Add the following i ngredi ent s to the jar in these proportions: 112 g. whole rye grains 2.0 g. Calcium carbonate (CaCO 3 ) .2 g. Potassium phosphate (K 2 HPO 4 ) (opt i onal ) 180 ml tap or distilled water (cf . Fig. 22-25) The potassium phosphate, if unavai l abl e, can be omi t t ed. The calcium carbonate need not be of great pur i t y: powdered oy- ster shell, powdered limestone, or powdered chalk is suitable. When the ingredients have been added to each jar in the proper proportions, the lids should be screwed loosely onto the jars, with the rubber seal of the inner lids inverted so that the jars will not seal during sterilization (Fig. 26). Now the jars containing the rye can be sterilized. Barely cover the bottom of the pressure cooker with tap water. Place the jars in the cooker, making sure that the lids are loose (Fig. 28). If one's pressure cooker is large enough to permit, jars can be stacked in two tiers without di f f i cul t y (Fig. 29 & 30). Seal the lid of the pressure cooker, but leave the stopcock open as before, until a head of steam begins to vent from the stopcock. Then close the stopcock, and bri ng to 15-20 Ibs. pressure. Reduce heat when this pressure is reached so that pressure is maintained but does not increase. This is about medium heat on an electric stove. Sterilize at this pressure for one hour. Remove from the heat and allow the pressure to return to zero before opening the stopcock. Open the stop- cock at this point and allow excess steam to escape; then re- move the lid from the pressure cooker. Remove one of t he jars, tighten the lid down finger-tight, and carefully examine the jar for cracks and immediately discard any flawed jars (Fig. 31) then shake the remaining jars vigorously (Fig. 31). One can inscribe the jars with the date of inoculation or other suitable code-number (Fig. 32) in order to keep track of the schedule of shaking the jars. One will note on removing the jar that the rye has absorbed the water and swelled to several times its previous volume. After shaking, leave the lids tight- ened until the jars have cooled. Place the jars into the pre- sterilized inoculating hood (if available) or onto the clean, disinfected working surface (Fig. 10). Then let the jars cool for at least two hours or to room t emperat ure. When the jars have cooled completely to room tempera- ture, and are no longer warm to the touch, they are ready to be i nocul at ed. This step is best accomplished using a dispos- able, presterilized #1 1 scalpel (Fig. 33). These scalpels can be obt ai ned from any medical supply house. Flame the blades of the scalpel over an alcohol lamp (Fig. 33). Insert the scalpel into an agar mycelial culture grown in a petri plate or baby food jar, and cut a grid into the surface, so that small squares of agar of about 1-1.5 sq. cm. are formed (Fig. 34). One can get about 9-20 small squares of agar from a single four-inch petri dish in this way (Fig. 35). Cultures grown in slant t ubes are obviously unsuitable for this.step, because of difficulty in removing squares of agar from the tubes. At each transfer re- sterilize the scalpel blade by flaming, insert into the agar cul- ture, and spear and remove one of the squares of mycel i um- covered agar (Fig. 36) and transfer rapidly to one of the ma- son jars, l i f t i ng t he lid of the jar just enough to insert the ino- culum (Figs. 37 & 38). Firmly tighten the lid and shake the jar vigorously to spread the inoculation points. Then be cer- tain to reloosen the lid; it must be left loose enough to allow di ffusi on of oxygen from the surrounding air, as proper aera- tion is essential for the f ungus to grow. Repeat t hi s inocula- tion step for each jar. An al t er nat i ve met hod of i nocul at i on was suggested to u s by a mycologist friend, as possibly effective in increasing 34 35 fig. 17 : Maki ng a box: cut t i ng a fig. 18: Appl yi ng silicone ma r i ne window in the lid . . . glue. . . fig. 19: Cl ear pl ast i c vi nyl i s gl ued fig. 20: Compl et ed box. over the window . . fig. 21: A set of j ar s t o be f i l l ed and fig. 22: Ma t e r i a l s for maki ng t h e sterilized. rye medi um. fig. 23: Addi ng weighed rye to t he fig. 24: Two gr ams of powder ed jar. oyst er shel l (CaCO 3 ) are added. fi g. 25: 180 mi l l i l i t e r s of wa t e r are fig. 26: Mason j ar lid wi t h r u b b e r added. edge up. fig. 27: The Ait-American 94H-2 fig. 28: Loading t he pressure cooker, pressure cooker. 36 37 fig. 29: The first l ayer of jars. fig. 30: The second l ayer of jars. fig. 3 1: St eri l i zed j ar s ar e checked fig. 32: Wr i t i ng the dat e on the j ar s, for cr acks, t hen shaken. fig. 33: F l a mi ng t he scalpel, fig. 34; Mycel i um on agar i s cut i nt o sect i ons. 38 t he st eri l i t y of the procedure and thus cut t i ng down contami- nat i on. This method is a st andard approach to fungal inocu- l at i ons in mycological work, and so far seems promising, al- though we have not investigated it thoroughly enough to know if it is t he answer to cont ami nat i on problems. This met hod is carried out as fol l ows: Sterilize a bi t of wat er by placing it in a flask sealed with al umi num foil and placing in the pressure cooker wi t h t he mason j ar s. When one is ready to inoculate, take up about 10 cc (ml ) of the wat er in a dis- posable, presterilized plastic syringe, and inject it ont o t he surface of an agar mycel i al cul t ur e (sl ant -t ube cul t ures are ideal for this). Reseal t he cul t ur e and swirl the water around vi gorousl y u n t i l fragment s of mycel i um become vi si bl e sus- pended i n t he water. Then again take up the wat er -mycel i al suspension in the syringe, and use the syri nge to i nocul at e each mason j ar by i nject i ng 1 or 2 ccs i nt o each j ar . As wi t h the scal pel , the needl e of the syringe can be flamed over an alcohol l amp before making each inoculation. Mycelial growth should become vi si bl e from one or more poi nt s in the j ar by 3-4 days after i nocul at i on. Af t er t he rye has been i nocul at ed, a period of wai t i ng and car ef ul observation follows. The j ar s should be mai nt ai ned at a roughly constant t emperat ure of 70-80 F., and 957c rela- tive humi di t y. Since t he lids r emai n on the jars t he humi di t y will tend to be high and need not be worried about. Mai nt ai n- ing the relatively high and const ant t emper at ur e, however , is i mport ant t o pr omot e ear l y and r api d mycelial growt h. The mycel i um gives off heat as it grows and a st yrofoam box of the sort used by t ropi cal fish wholesalers is i deal f or holding t hi s heat and t hus self-incubating the jars. Duri ng t he first three days af t er i nocul at i on, the mycel i um wi l l grow off t he agar i nocul um and ont o the rye. By the f our t h day, the my- cel i um will have grown r a d i a l l y out ward from the i nocul um in al l di r ect i ons to form a mat of growth slightly smaller t han a fi ft y-cent pi ece. When mycelial growth has reached t hi s stage, f i r ml y tighten the lid and again shake t he c u l t u r e vigorously, to br eak up the mycelium and redi st ri but e the i nocul um t hr oughout t he rye. During the shaking process you wi l l r un across ob- viously cont ami nat ed jar s; s i mpl y remove t hem and set them 40 aside for washing out later. Reloosen the lid i mmedi at el y af- ter shaking and al l ow the c ul t ur e to st and for 34 days. At t he end of this t i me, if the lids are loose enough to permi t proper aer at i on, myc e l i a l growth from many di fferent points in t he rye should be apparent (F i g. 39). If the rye has not been compl et el y per meat ed by t he mycel i um at the end of the si xt h to ei ght h day aft er i nocul at i on, the cultures can be shaken a second t i me. Compl et e permeation of the rye should be observed anywhere from the eighth to the f our t eent h day. At this time, the rye should be compl et el y permeat ed by the snow-white mycel i um, which may occasionally be lightly tinged wi t h blue. If growt h of any ot her color is observed, or if the rye is only partially permeat ed, then the culture is con- t ami nat ed and should be di scarded. It should be noted t hat these time paramet ers given f or compl et e per meat i on of t he rye by the mycelium can var y wi del y. I n some cases, compl et e per meat i on can take place i n under si x days: in ot hers, two or even t hr ee weeks are re- qui red. Exact l y what f act or s govern per meat i on time are not clear, but we suspect that some external envi ronment al para- meter, such as t emper at ur e var i abi l i t y, humi di t y or partial pressure of oxygen, may be r esponsi bl e. Int erest i ngl y, we have found t hat mycelial growt h seems to occur more vigorously in c ul t ur e s grown in the war mer , coastal part s of t he count r y (California) while mycel i um grown in high-altitude, low-hu- midity regions (Col orado) seems to be sl owed by some sort of external envi r onment al stress, possibly al t i t ude. Even if some unc ont r ol l a bl e envi r onment al fact or is responsi bl e for the re- tarded growth exhi bi t ed by the mycel i um in cer t ai n localities, t hi s need not di scour age those a t t e mp t i n g t o work with t h e organism in these areas. All t hat is r equi r ed is somewhat more pat i ence. Periodic shaking of t he cul t ur es can hel p, as it seems to pr omot e growt h by f aci l i t at i ng aeration. The pr obl em can also be combated by using only vigorously growing mycelium grown on a nut r i e nt -r i c h agar medi um. The mal t ext r act agar recipe is in this respect pr ef er abl e to PDA. If t he rye grai n medi um i s also enriched by t he a d d i t i o n of ma l t ext r act a nd yeast ext r act , and vi gor ous cul t ur es grown on a rich medi um are empl oyed, permeation t i me even under condi t i ons of en- vi r onment al stress can be cut to t he nei ghbor hood of 10-14 41 days. It is qui t e i mpor t ant t hat the mycelium permeat es t he rye in t he mi ni mum time possi bl e, because t he sooner the fungus takes over the medi um compl et el y, t he less chance is there for ot her cont ami nant s to gain a foothold. When one or more jars of rye have been completely per- meated by mycel i um, t he third step in the procedure is com- pleted and one is ready to move on to t he f our t h step, casing. Before discussing this step, however, it is perhaps advisable to insert a word of caut i on with respect to the third step. This step, getting the mycelium to grow out and permeate t he rye, seems to be the most di f f i cul t and di scouragi ng step in the whole procedure. The peculiar headaches t hat one is faced wi t h in t hi s step can be summed up in one wor d: cont ami na- tion. For some reason, cont ami nat i on seems to be a much more serious problem at this stage t han at t he stage of grow- ing on agar, probabl y because rye is a sui t abl e medi um for such a great variety of lower organisms. Anyone at t empt i ng this step, in fact, is almost sure to receive a real education in the number of fungal and bacterial "weeds" t hat exi st to plague the a ma t e ur mycologist. Our experi ence has been t hat two cont ami nant s in par t i cul ar are qu i t e persistent and seem- i ngl y impossible to ent i rel y el i mi nat e. One is a crust y, rapidly- growi ng blue-green mold with a medi ci nal odor, probably a Penicilliitm or Aspergillim. The other is an uni dent i f i ed bac- teria t hat exudes a yellowish slime ont o t he side of the j ar and t hat smells strongly of rotten apples. Spores of bot h of these organisms must be so commonl y present in nature that they manage to cont ami nat e some of the cul t ur es despite the most careful inoculation procedures. The mol d shows up ra- pidly and can be qui ckl y spotted. An y cul t ur e seen to have this cont ami nant can be considered a loss. The bacteria takes longer to become obvious but wi t h practice one can learn to spot it wi t hi n a few days after i nocul at i on. Cul t ures contami- nat ed wi t h this organism are also almost impossible to salvage. The bact eri a is anaerobi c, that is, it can grow in t he absence of oxygen. Our experience is that it seems i ndi f f er ent to the presence or absence of oxygen, and grows in ei t her si t uat i on. The mushroom mycel i um is qui t e aerobic, in fact proper aera- t i on is essential for its growth; therefore proper aeration can 42 afford t he f ungus something of a competitive advantage against this organi sm. Other cont ami nant s will occasionally be seen, although not with the regularity of t he two already ment i oned. These may include black, olive green or sulphur-colored molds, and sometimes a dirty-grey rapidly growing mol d that is probably a Neurospom. Contaminants of any kind are not good and it is advis- able to discard immediately any culture seen to be contami- nated. Although the mycelium can co-exist with some of the slower-growing fungal cont ami nant s, it is still best to discard any contaminated cul t ures in order to avoid spreadi ng t he plague. The cleaning of jars should be done as far away from the inoculation area as possible and should be done by some- one who is not involved in making sterile inoculations. Jars that have been contaminated should be washed in a strong solution of Clorox and water before being reused. The best way to deal with cont ami nat i on is to not allow it to become established in the first pl ace, by being extremely meticulous about one's sterilization and inoculation procedures. Always make sure that the jars are sterilized with wet heat , not dry, by allowing a head of steam to build up in the pressure cooker before closing the stopcock. An i nocul at i ng hood, even if it is as simple as a cardboard box with clear plastic in one side, be- comes almost i ndi spensabl e at this stage. Al ways use sterile, uncont ami nat ed agar cultures as t he source of i nocul at i on. Make sure that the working surface, and the inside of the ino- culating hood, are thoroughly disinfected before inoculation. Also make sure that one's hands and arms are clean before in- oculating, and wear l at ex rubber gloves if possible. Spray your gloved hands with spray Lysol before reaching into the hood. Use disposable presterilized scalpels for maki ng inoculations, or if the i mpl ement employed is to be reused, be cert ai n that it is absolutely clean for each batch to be inoculated. It does not hur t to swab it with alcohol beforehand. Be certain to fl ame t he implement thoroughly in an alcohol fl ame before making each transfer. Practice making t ransfers as r api dl y as possible, so that nei t her the receptacle containing the inocu- lum, or the mason jar are kept open longer t han necessary. 43 Finally, be abs ol ut el y ruthless in discarding contaminated cul- tures. Not hi ng less t han complete permeation of t he rye by t he snow-white mycel i um should be consi dered acceptable. If these procedures are followed rigorously, some c u l t u r e s will undoubt edl y still succumb t o c ont a mi na t i on, but t he number can be held to a mi n i mu m. 44 STEP IV: CASING When one or more j ar s have been compl et el y per meat ed by myc e l i u m, one can move on t o t he fourt h step i n t he pro- cess t hat l eads di rect l y t o t h e pr oduct i on of mu s h r o o ms . In commerci al mushroom c u l t u r e , t hi s step is cal l ed casing. In the met hod out l i ned here, casing consi st s of removing t he dome and band lid of t he j ar , and covering the surface of t he per meat ed rye with about V i to % in. (V i c up for quar t j a r s ) of sterilized soil (F ig. 40 & 41). The soil can be appl i ed whi l e dry, b u t t he n shoul d immediately be shaken level, t he n wet t ed to fi el d capacity usi ng chlorine-free tap or distilled wat er ap- plied in a fi ne-mi st spray (Fig. 42 & 43). A f i ne-mi s t spray must be used to avoid seal i ng t he sur f ace of t he casing s oi l . Field capaci t y can be gauged by t he following rule of t humb: spray t he casing soil j us t enough so t hat t he soil is moi st ened t hroughout , but no wat er passes through t he soil i n t o t h e my- c e l i u m. In ot her wor ds, moi st en t hor oughl y, but do not sa- turate, t he soil. Aft er appl yi ng and moistening the casing soil, di scar d t he l i ds and mai nt ai n t he c u l t u r e s i n a hi gh humi di t y envi r onment . A large styrofoam cooler wi t h a wi ndow c u t i n t o t he l i d and covered with clear or translucent pol yet hyl ene is excel l ent for this (Fig. 17-20). Spray the cultures daily with a fine-mist spray j ust enough to make up for moi s t ur e lost t hr ough evap- orat i on (F i g. 44). Each cased j a r requi res 2-3 good squi r t s of wat er per day to maintain cont i nuous f r ui t i ng. Do not exceed field capaci t y. A good t est f or proper moi s t ur e cont ent is t hat the sur f ace of t he soi l shoul d f eel moist and spongy to t he touch. Boxes of newl y cased jars should be stored in chrono- logical or der . Dur i ng t he next t wo t o t hr ee weeks, t he mycel i um wi l l begin t o grow up into t he easing soil, p e n e t r a t i n g i t t o j u s t beneat h t he s ur f a c e (F i g. 45). The myc e l i um may occasion- al l y break out ont o t h e t op of t he soil and begin t o spread across its surface, and par t of t he purpose of spr ayi ng dai l y is t o keep t hi s surface growt h ' ' knocked down" wi t h t he spray (Fig. 46) . If you are under -wat er i ng your jars t h i s surface my- 45 47 celium wi l l turn bl ue with t hi r st . As the mycelium grows into the casing soil.it will begi n to form a net wor k of ropy strands visible at t he i nt er f ace of the soil and the glass. This ne t wor k gradual l y gains more and more i nt er sect i ng nodes, and by the 14th to 20th day a f t e r casing, these nodes have di fferent i at ed i nt o t i ny wh i t e dots distributed t hrough t he casing soil along t he peri met er of t he j ar . These dots are t he young mushroom pr i mor di a; gradual l y they enlarge and i ncorporat e more my- celia, slowly t a ki ng on t he appear ance of t i n y mushrooms wi t h s qua t , fat st ems a n d dar k browni sh heads. Thi s i s t he "pin" stage in t h e devel opment of the mushroom and t he pri mordi a at this stage are about 1-2 mm. in l engt h. These pins cont i nue to enl arge and some wi l l begin to t hrust above the surface of the casing s oi l , both at the sides and in t he cen- ter of t h e jar. Once the young mushrooms have penet r at ed t he s ur f ace of t he casi ng soil, anot her f i ve t o t en days i s re- qui red for t hem t o r each f ul l ma t ur i t y. At ma t ur i t y t h e mushrooms wi l l be several i nches tall and may have caps rang- ing from one to several cm in di amet er (Fig. 47 & 48. a nd color phot os). We have also f ound t h a t these mushr ooms do respond favorabl y to l i ght , and t hat a daily 10-hour Grow- l ux cycle resul t s i n mushrooms wi t h l arger caps and short er stems than t hose grown wi t hout any special l i ght i ng. Prob- ably it is not advi sabl e, however , to place t he cul t ures in di r e c t sunlight for prolonged periods. While many mus hr ooms will grow to f ul l size, appr oxi mat el y an equal number wi l l grow to about hal f-si ze or less and t hen cease to gr ow; these ''aborts" can still be pl ucked, dr i ed, and used, although they are not as aest het i cal l y appealing as t he f ul l y ma t ur e d specimens. Wi t h pract i ce one can learn t o spot aborts ear l y and remove t hem from t he cul t ur es. Abort ed mushrooms l e f t i n t he c u l t u r e s are susceptible t o at t ack by bact eri a whi ch qui ckl y render t hem bot h ugly and unusabl e. Occasionally mushr oom primordia will form down toward t he bot t om of t he j a r and grow to mat ur i t y b u t wi l l not br eak t he sur f ace of t he casi ng soil. It i s possible t o i n h i b i t t h i s effect somewhat by wrapping j ar s wi t h t i n f o i l t o t he top of t he cas i ng soil (F i g. 45). If t hi s be- gins to happen to a great e xt e nt , it is possible to c a r e f ul l y break t he j a r and remove t he myc e l i um in a single block. Then pr i mor di a al ong t he si des of t he bl oc k, whi ch would otherwise abort due t o cons t r i ct i on by t he j ar , wi l l be abl e t o 48 reach mat ur i t y. The j a r mus t be broken c a r e f u l l y, however, t o avoi d br ui s i ng t he f ungus and injuring onesel f . Af t e r t he mycel i um has been broken out , one must also make sure that it is not al l owed to become too dr y. A vari et y of t ypes of easing soils have been f ound to effect i vel y promote f r u i t i n g . We have found t he following mi xt ur e to be one of t he best: 2.5 liters peat moss 3.5 liters fine vermiculite 4 liters washed fine sand 2 l i t er s cal ci um carbonate (f i nel y crushed oyster shel l ) Powdered oyster shell is sold as a feed suppl ement by many feed compani es. We have also f o u n d t hat a mi xt ur e of one part Mi ca-peat (50/50 vermiculite-peatmoss mi xt u r e ) t o one par t pot t i ng soi l will work, and even unadul t er at ed r i c h gar- den l oam has been f ound sui t abl e, t hough its unst eri l e condi- t i on makes c ont a mi na t i on a pos s i bi l i t y. There is room for f u r t h e r experi ment at i on with other types of casing mi xt ur es : one mi ght t ry casing wi t h finely granulated horse-dung or cow-dung, or a mi xt ur e of horse- or cow-dung and f i n e l y chopped wheat-straw. Casing wi t h l eaf- mold mul ch mi ght also ef f ect i vel y promot e f r u i t i n g , and i n fact might encourage the f r ui t i ng of smaller and more delicate species of Psilocybe t hat do not seem abl e to f r ui t when t he mi xt ur e given above is used. The object is to find a casing soil t hat i s por ous enough t o al l ow air t o r each t he mycel i um, and t ha t at t he same time is l i ght enough to allow t he young mushrooms t o penet r at e easily t hr ough t he surface. Steriliza- t i on of casing soil i s us ua l l y recommended but we f ound i t unnecessary when relatively st eri l e commerci al l y bagged ma- t er i al s were used, i f you wish you may s t e r i l i ze casi ng soil be- fore use at 15-20 Ibs. pressure for one hour . It can be steri- lized dr y, or wet t ed f i r s t t o f i e l d capaci t y. Casing soil can be stored i ndef i ni t el y in a t i ght l y sealed large glass j a r . or in a pol yet hyl ene garbage bag. If a glass jar is used, it can be steri- lized wi t h t he casing soil in i t . 49 STEP V: HARVESTING, PRESERVING, AND DOSAGE As the mushroom mat ur es to full size, the cap will en- large and become more globular in shape. The gills will at first be covered by a flap of tissue, called the veil, that connects the margin of the cap with the stipe. As the cap enlarges, this veil will det ach from t he cap margin to form an annul us , or ring of ruptured veil tissue, on the stem (cf. color figures). The mushroom can be harvested as soon as veil rupt ure has occurred. If a spore print is to be collected, however, t he mushroom should be allowed to flatten out to an umbrel l a- shape before harvesting. Freshly harvested mushrooms can be eaten fresh, or dried and stored. We recommend that the mushrooms be dried in a gas oven at low heat (140 F. or less) for 6-10 hours (Figs. 49 & 50). They may also be dried under a heat - lamp, or placed on a screen and dried over a heat i ng vent. Mushrooms are f ul l y dr i ed when hard to t he t ouc h, like crackers, with no spongy feel at all (Fig. 51). To preserve maxi mum pot ency, dried mushrooms should t hen be sealed in five gram increments in plastic bags from which the air has been wi t hdrawn (Figs. 52-54), and this in turn placed in a tightly sealed glass vessel or other moisture-proof container, and frozen. Dried mushrooms left in the open air quickly lose their potency. Fresh mushrooms should not be frozen with- out drying first, as freezing them in this condition will turn them i nt o a black, gooey mess. Fresli mushrooms can, how- ever, be preserved in a pl ast i c bag in the vegetable pan of the refrigerator for about a week to ten days. Fresh mushr ooms older t han this should either be eat en or dried to prevent spoilage. The dried mushrooms cont ai n from .2 to .4 percent of psilocybin (Schultes, et al., 1973) by wei ght . Some strains of Stropliaria cubensis have been reported to cont ai n as much as .5% psilocybin (Wasson & Hei m, 1959, p. 260). Psilocin is present onl y in trace amount s . A dose of about 10-12 mi l l i - grams of psilocybin, or about 5 g. dr y wei ght of mushrooms, or 50 g. wet weight, is suffi ci ent to manifest the full spect rum 51 fi g. 48; Grasp t he st al ks f i r ml y fi g. 49: Remove t he soi l from t h e when harvesting. bot t oms of the s t ei ns . _ MBaMM^weMMBBnHI fig. 52: They are pl aced in pl as t i c fig. 53: Bags are heat-se; bags. bag seal er. 52 fig. 54: F resh mushrooms and dried ones r eady for t r e e / i n g . 53 of hallucinogenic effect s in a 160 Ib. adul t . These effect s in- clude visual and audi t ory hallucinations, ext reme hilarity, dis- tortions of time and space perception, and a sense of emotion- al detachment from the environment. Less mar ked effects can be detected at doses as low as 4 mg., which is about 2-3 dried mushrooms. Fresh mushrooms seem to be somewhat stronger than dried ones. Psilocybin is one of the least toxic of all hal- lucinogens. While a full effective dose is 10 mg., its LD 50 (lethal dose for 50% of the sample) in the mouse is 280 mg./kg. of body weight. (Schultes & Hof mann, 1973). Mesca- l i ne. by comparison, has a minimum effect i ve dose of 200 mg. for an average-size adult, and a toxicity 2.5 times that of psi- locybin (Aboul-Enein, 1974). The state of mind induced by a full dose of mushrooms is one of euphoria and calm lucidity, with no loss of coher- ency or clarity of thought. The hallucinations seen with the eyes closed are colorful, hard-edged, and highly art i cul at ed, and may range from abst ract geometrical forms to visions of fantastic landscapes and architectural vistas. These hallucina- tions are most intense when the mushroom is taken in the set- ting preferred by the Mazatecans; inside at night in complete darkness. On the other hand, if one is in a nat ur al setting and di rect s the focus of the senses outward to the envi ronment , one discovers t hat one's senses seem keyed to their highest pitch of r ecept i vi t y, and finds oneself hearing, smelling and seeing things wi t h a clarity and sensitivity seldom, if ever, experienced before. Al t hough it should be clear to anyone who has read this far that cultivating these mushrooms is an endeavor requiring time, pat i ence, care and humi l i t y, and one fraught with i t s own peculiar probl ems, once one has par t aken of these wonderful gifts of nature and experiences the exalted consciousness that they can bring about , we t hi nk t hat they will agree with us that t he effort i nvol ved provides its own ample reward. AFTERWORD Approximately sixty days after you began the isolation of spores the first harvest will be possible from your rye-filled jars. Mushroom growing is l i ke alchemy in t hat there is a divi- sion of t he work i nt o practical effort and visionary reward. The organic psilocybin within the mushroom is qual i t y con- trolled by the very stable and ancient genes of the Stropharia. You, as the propagator and spiritual friend of the mushroom can form a deep relationship with t he mycelial ally and agai n and again make far j our neys i nt o i t s visionary realms if you observe a few simple rules. Tolerance to psilocybin is easily acquired if trips are taken more often than once a week. If one does acquire a tolerance it can be gone around by either upping the dose or by laying off for a couple of weeks to al- low your body to recover its equilibrium. We recommend the latter course even though the t oxi ci t y of psilocybin is so low that raising the dose is a valid alternate course. Now our little handbook closes. Advice from this poi nt on can only come in general i t i es. Take the mushrooms that you have gr own, take them in the darkness as the Indi ans of Mexico who have used them for centuries do. Smoke a lot of your favorite hash to synergize the behind-the-eyelids hallu- cinations and prolong t hem. Psilocybin is light shedding illu- mi nat i on on a landscape both within and wi t hout t he mi nd and body of man and previously invisible. The expl or at i on of this vast region by persons whose mental equipage is that of the modern West has only begun. Only a moment has passed since our culture has rediscovered, through the work of Was- son and others, the ancient and unpl umbed relationship be- tween the vision-causing mushrooms and our own strangely gifted species. You are a pioneer in a world whose fut ure is undet er mi ned and whose living organisms are full of singular- ities and surreal t r ansf or mi ng promise. 54 55 CONVERSION TABLE This conversion table is included for those who may lack scales or ot her equi pment to make accurat e measurement s of requi red ingredients. Although one should always try to mea- sure the i ngr e di e nt s as accur at el y as possible, and the purchase of an inexpensive scale and graduated cylinder is well worth whi l e, this table can be u s e f u l i n appr oxi mat i ng meas ur ement s and cal cul at i ng vol umes if such equipment is unavailable. 112 g. r ye is just over y 2 cup 2 g. powdered oyster shell is a l evel V 2 teaspoon 180ml . H 2 0i s 6f l . oz. 1 g. yeast ext ract is a level V 2 teaspoon 1 g. Difco agar is 1 sl i ght l y packed level V 2 t easpoon [ liter (1000 ml. ) - ] quar t (appr ox. ) 1 baggie (Glad t ype) wei ghs 1. 5 g. 56 A CHRONOLOGY OF PS1LOCYBIAN* MUSHROOMS Compiled by Irimias the Obscure 300-500 B.C. In t he l at t er hal f of this c e n t u r y, "mushroom stones" wer e f ound in highland Gua t e ma l a dating back at least as f ar as 300- 500 B.C. c. 300 A. D. F rescoes have been f ound in c e nt r a l Mexico wi t h mush- room designs indicating the existence of a mushroom cult at this time. 1502 A.D. Spanish conqui st adores observed psilocybian mushrooms bei ng served at t he cor onat i on feast of Moct ezuma. 1547-1569 F r a y Bernadi no de Sahugun, a Spanish cl eri c, wrot e His- toria de las Cosas de Nueva Espauia (also known as thsFlorentine Codex) which refers to "nanacatl" (= teonanacatl = flesh of the gods = psilocybian mushroom). Sahugun st at es t hat t he mushrooms "are h a r mf u l and i nt oxi cat e like wine." F u r t h e r , those who i ndul ge "see visions, feel a f ai nt ness of hear t and ar e provoked to lust." 1651 Dr. Francisco Her nandez, a Spanish physician studying Ce nt r a l Amer i can Indi an her bal medi ci ne r epor t ed t h r e e t ypes of mushrooms which wer e worshi pped by Mexican nat i ves. He report ed t hat the in- gestion of t hese caused "not deat h but a madness that on occasion is l ast i ng, of whi ch t he sympt om is a ki nd of uncont r ol l ed l aught er . . . these are deep yellow, acrid, and of a not displeasing freshness. There ar e ot hers again, whi ch wi t hout i nduci ng l a ught e r , br i ng bef or e t he eye all s or t s of t hi ngs, such as war s and the likeness of demons. Yet ot her s t here are not less desi red by pri nces for t hei r festivals and b a n - quet s , and t hese fetch a high pri ce. Wi t h night-long vigils are they s o u g h t , awesome and t e r r i f yi n g . This kind i s t a wn y and somewhat acr i d/' 1906 Stropharia cubensis is described by Carle in a Cuban agronomy j o u r n a l . 1914 A. F . Mer r i l l of Yale Uni ver si t y published a paper in Science de- s cr i bi ng t he hal l uci nogeni c e f f e c t s of i ngest i ng Panaeolus papillona- ccus from Oxford County, Maine. Although the identification of the mu s h r o o m may he i n e r r or , t he e f f e c t s descri bed are ve r y p r o b a b l y due t o psi l ocybi n and psi l oci n. F u r t h e r , t he a r t i c l e descr i bes d i f f e r - ent r eact i ons t o t hi s hallucinogenic mushroom whi c h i s c ompa r e d t o hashi sh and peyot l i n the t ext . '"Psilocyhian" i n t hi s cont ext means any mushr oom cont ai ni ng psi l ocybi n. 57 19 15 Ameri can bot ani st Wi l l i am E. Safford a t t e mp t e d t o i dent i fy t he t eonanacat l of the Aztecs. He claimed t hat sacred mushrooms had never exi s t ed, and t ha t t he t eonanacat l r ef er r ed t o by the 16th cen- t u r y Spanish chroniclers were act ual l y dri ed peyotl but t ons . Safford's t heor y was wi del y accept ed by t he s c i e nt i f i c c ommuni t y for the next t hree decades. 19 19 Dr . Bias P. Reko, who had carried out ext ensi ve ant hropol ogi cal and bot a ni c a l wor k i n Mexico for mor e t han 25 year s, publ i shed an art i cl e i n a Mexi can ant hr opol ogi cal j our na l st at i ng t h a t nanacat l (= t eonanacat l ) was a hal l uci nogeni c mushroom. However, some of Rcko's earl i er work had been in error and this report was di s count ed. 1923 In a l e t t e r to t he U.S. Nat i onal Mus eum, Dr . Reko st at ed that t e ona na c a t l "is act ual l y, as Sahugim st at es, a fungus whi ch grows on dung heaps and which is still used under t he same old name by the Indi ans of t he Sierra Juar ez in Oaxaca in t hei r religious feast s. " 1936 Vi ct or A. Reko (B.P."s brot her) publishes Magische Gifte. In i t . he wrongly suggests that t eonanacat l might be a species ofAmanita. 1936 Ing. Rober t o J. Weitlaner obtained some t eonanacat l in Oaxaca. He was the first white man in modern times to have done so. He sent t he specimens to B.P. Reko, who sent them to Ha r va r d, where they arrived in a decomposed state and t hus escaped i dent i f i cat i on. 1938 VVe i t l a n e r ' s daught er, along with ant hr opol ogi st Jean Basset Johnson and two others a t t e nde d a mushroom rite in Huat l a, Oaxaca. These were the first wh i t e s to at t end a mushroom ceremony. 1938 Har var d bot a ni s t R. E. Sc l i ul t e s t ravel l ed t o Oaxaca and obt ai ned from native i nf or mant s two specimens of t wo di f f e r e nt gener a: Pan- aeolun campanulaius var. sphinctrinus, 'and Stropharia cubensis. In his field not es he descri bed a t hi r d specimen: Psilocybe caeruiescem var. Mazateconiin. 1953 R. Gordon Wasson and his wife Valentina became aware of the exi s t ence of a mushr oom cul t in cent r al Mexico. This ambi t i ous couple set out to prove the t heory t hat religion came di rect l y from t he use of hal l uci nogeni c pl ant s . The Wassons t r avel ed to Mexi co and were gui ded by Ing. Rober t o J. Wei t l aner t o t he mount ai nous vi l l age of Huatla de Jiminez in Oaxaca. 1955 R.G. Wasson and Al an Ri char dson became the first two Ameri- cans to a t t e n d a mushroom r i t ual and ingest the mushrooms. The mushrooms were t aken under the supervi si on of Mar i a Sabi na. a local c u r a n d e r a . By 1957, news of t h i s r i t ua l had reached the world through art i cl es in several popul ar magazi nes and t he Wasson's book, Mushrooms, Russia, and History. 58 1956 Wasson invited Roger Heim, a F rench mycologist to Oaxaca to research t he use of t he sacred mushrooms. Heim identified fourt een species and several subspecies belonging to three gener a, Psilocybe, Stropharia. and Conocybe. Several of these species were new to my- cology, but had been utilized as hallucinogens by the nat i ves for cen- t uri es. 1957 Mycologist Dr. Rol f Singer and two young Mexi can bot ani st s, M.A. Palacios and Gaston Guzman, arrived in Oaxaca to do t axo- nomic work on the mushrooms. 1958 Dr. Al ber t Hof mann. a Sandoz chemist from Basel, Swi t zerl and, isolated two active agent s and named them psilocybin and psilocin a f t e r t he genus Psilocybe. 1960 While vacat i oni ng in Cuernavaca. Mexico, Har var d psychologist Timothy Leary ate a dose of the mushrooms. La t e r , he wr ot e: ". . . it was the classic visionary voyage and I came back a changed man . . . You are never the same after you've had t hat one flash glimpse down t he cel l ul ar time t unnel . You are never the same af t er you've had t he veil drawn." 1960 Dr. Leary and an associate, Dr . Ri chard Al per t , obt ai ned a sup- ply of synt het i c psilocybin f r om Sandoz for use in an experi ment with prisoners in Concord St a t e Pri son, Massachuset t s. Initial results wer e very promi si ng, pr i soner s released following an experi ence wi t h psilocybin seemed less likely to be r ear r est ed and r et ur ned for parole violations t han other parolees. 1960 Aldous Huxley ingested 10 mg. psilocybin in a group under the supervision of Ti mot hy Leary. Huxl ey "sat in cont empl at i ve calm t hr oughout ; occasionally produced relevant epigrams; r epor t ed the experi ence as an edifying philosophic exper i ence/' c. 1965-66 Laws against the sal e, manuf act ur e, and possession of LSD, mescaline and psilocybin are passed by paranoid legislatures af t er be- ing per suaded by a hysterical press. The New Yor k St at e l egi sl at ure def er r ed hearings on one bill to out l aw hallucinogens unt i l a f t e r t he law was vot ed on and passed! 1966 By this time several illicit labs were set up to manuf act ur e hal l u- ci nogeni c dr ugs in response to the growing demand by users. 1967 React i ng to e r r one ous tales of massive chromosome damage pro- duced by LSD use, users began to demand organic drugs such as psi- l ocybi n and mescaline. Compared to LSD, hal l uci nogens such as these are r el at i vel y expensi ve t o ma nuf a c t ur e . Many unscrupul ous deal er s sold LSD as psilocybin. Most of t he tabbed or capsuled psilo- cybi n on the st reet from 1967-75 was act ual l y LSD, or LSD cut with PCP. 59 1970 A Key to the North American Psilocybin Mushroom was pub- lished by Leonard Enos in Cal i forni a. This poorly-illustrated but well- written gui de i ns t r uct ed laymen where, wh e n and how t hey could obt ai n psi l ocybi an mus hr ooms i n n a t u r e . The book al so cont ai ned i nst r uct i ons for cul t i vat i ng mycel i um on agar. 197 1 Due t o popul ar demand for or gani c dr ugs , u n s c r u p u l o u s de a l e r s began lacing commercial mushrooms with LSD and selling t hem as psilocybian mus hr ooms . These spuri ous psi l ocybi an mus hr ooms have appear ed on t he st r eet d r u g ma r k e t as l at e as 1975 and can be d i f f e r - ent i at ed f r om most psilocybian mushrooms i n l l i a i t h e y ( a ) d o n ' t bl ue , and (b) t hei r ef f ect s last much l onger t h a n t he 4 l o 7 hour s char act er i s t i c of psi l ocybi n. 1975 The first l i vi ng c u l t u r e s of Stropharia cubensis were seen in lim- i t ed n u mb e r s on t h e under gr ound mar ket . 1975 Oss and Oeric (t hi s vol ume) br a ve l y ri sked r i di cul e to become t he fi rst to suggest t he ext r at er r est r i al origin of Stropharia cubensis. 1976 Technol ogy developed by t he a ut hor s (Oss & Oeri c. 1976) i s un- l eashed upon t he wor l d. The i l l i ci t hal l uci nogen t r a de cr umbl es be- cause of decent r al i zat i on br ought on by epi demi c of home Stropharia cul t i vat i on. Invasi on of Nor t h Amer i ca by ha l l uc i noge ni c mushrooms c ont i nue s , l eadi ng s hor t l y t o met amor phosi s of manki nd i n t o sym- bi ot i c species. BIBLIOGRAPHY Aboul -Enei n, Hassan Y.: "Psilocybin: a Pharmacological Profi l e," Am J Pharm, May-June 1974. Benedi ct , R.G., Tyl e r . V. E. . & Wat l i ng, R.: "Bl uei ng in Conocybe, Psi- locybe and a Stropharia Species and t he Det ect i on of Psilocybin," LloyJia, v. 30, #2, June 1967. Ca t a l f omo, P. & Tyl er. V. E. : "The Pr oduct i on of Psi l ocybi n in Su b - merged Cu l t u r e by Psilocybe cubensis, " Uoydia, v. 27, #1, March 1964. Enos, Leonard: A Key to the North American Psilocybin Mushroom, copyri ght 1970, Leonard Enos, Youniverse Pr oduct i ons. Mi l l e r . Orson K., Jr . : Mushrooms of North America, E.P. Du t t o n &Co. , New Yor k, second printing, no dat e. Pollock, Steven: "Psilocybin Mushroom Pandemic," J of Psychedelic Drugs, v. 7, #|, J a n-Ma r 1975. San Ant oni o, J.P.: "A La bor a t or y Met hod to Obt ai n F r ui t f r om Cased Gr ai n Spawn of the Cul t i vat ed Mushroom. Agaricus bisporus. " My- cologia, v. 63, 1971, p. 16ff. Scagel, Rober t F . . et al.: An Evolutionary Survey of the Plant Kingdom, f o u r t l i pr i nt i ng, 1967, Wadsworth Publishing Company. Inc. . Bel- mont. Cal i forni a. Schultes, R.E. & Hof mann, Al ber t : The Botany and Chemistry of Hal- lucinogens, 1973, Charl es C. Thomas, Publ i sher , Spri ngfi el d, 111. Si nger, Rol f: "Mycological Investigations on Teonanacat l , the Mexi can Hal l uci nogeni c Mushr oom, Pt. ]." Mycologia, v. 50, 1958, p. 239ff. Wasson, R. G. : "Seeking the Magi c Mus hr oom; ' Life Magazi ne, May 13. 1957. __ ; "The Divine Mushroom: Primitive Religion and Hal l uci nat or y Agent s. " Proc Am Phil Soc, Phi l adel phi a, v. 102, ^3, June 24, 1958. : "The Hallucinogenic Mus hr ooms of Mexi co: An Ad v e n t u r e in Ethnomycological Expl or at i on, " Trans A' Y Acad Sci, Ser. I I . v. 21. #4, F eb. 1959. & Wasson, V. P. : Mushrooms, Russia and History. New York, Pant heon Books. 1957. Out of p r i n t . & Hei m. R. : Les Champignons Hallucinogenes du Mexi que: Etudes Ethnologiques,Taxinomiques, Biologiques, Physiologiques et Chimiques. Pari s: Museum Nat i onal d'Historie Nat ur el l e. 1959. 60 61 GLOSSARY adnate (of gi l l s) - a t t a c h e d di rect l y to st al k. adnexed (of gills) - notched just at stalk. aerobic r equi r i ng oxygen in order to live. Opposite of anaerobic. annul us ri ng shaped remai ns of the par t i al veil whi ch hangs on the stalk. basidium (pi. basidia) cell on which the spores of a Basidiomycete are borne. basidiocarp basi di um-bear i ng st r uct ur e or "frui t i ng body" of a Basidio- mycete. Sometimes also called a carpophore. basidiospore - spore formed exogenously on a bas i di um, gener al l y fol- lowing karyogamy and meiosis. di karyot i c a fungal hypha havi ng two nucl ei per c e l l . diploid havi ng a single set of paired chromosomes (twice the numbe r of chromosomes as in the gamet es); 2n. Cf. haploid, havi ng only one full set of unpaired chromosomes; n. genome the basic set of chromosomes (n) cont r i but ed by each par ent . genus (pi . genera) - the major subdivision of a family or subf ami l y of pl ant s or ani mal s; it usually consi st s of more t han one species. heterothallic In Basidiomycetes, having thalli separable into two or more morphol ogi cal l y similar sexual st rai ns, wi t h conjugat i on occur- ring only when c ompa t i bl e ma t i ng types are paired. hypha (pi. hyphae) - one of the t ubul ar f i l ament s composing mycelium. indole - a whi t e cryst al l i ne compound, Cs H 2 N, having the same het ero- cyclic fused ri ng s t r uc t ur e as the ammo acid tryptophane. The indole st r uct ur e is i ncor por at ed into the s t r uct ur es of many hal l uci nogeni c compounds. inoculation to implant mi cr oor gani sms or f ungal mycel i um i nt o a cul- t ur e medi um. The mycel i um used for t hi s is called t he inoculum (pi. inocula). inoculating loop i mpl ement used for making i nocul at i ons, consisting of a long handl e wi t h a length of stainless steel or pl at i num wire a t - t ached to the end and usual l y bent i nt o a loop at the t i p . karyogamy fusi on of the two nucl ei of the dikaryotic mycel i um. In the Basi di omycet es, karyogamy const i t ut es the only di pl oi d (2n) stage in the life cycle. meiosis r educt i on di vi si on of a cell in which the number of chromo- 62 somes is r educed f r om t he diploid (2n) to the haploid (n) state. Meiosis pr oduces sexual cells or gametes. milliliter (abbr. ml.) one-t housandt h part of a liter. 1 ml. = 1 cubic cent i met er (cc) in vol ume. monokaryotic hyphal condition in which the cells cont ai n a single haploid nucl eus ; e.g., t he primary mycelium of Basidiomycetes. mycelium (pi. mycelia) t he veget at i ve body of certain complex fungi , consi st i ng of an aggregation of hyphae. phylogeny the evol ut i onary devel opment of a species of pl ant or ani - mal. somatogamy fusion of somatic (body) cel l s r at her t h a n di f f er ent i at ed sexual cel l s as in the Basidiomycetes; does not i ncl ude karyogamy. species the major subdivision of a genus or subgenus; it is composed of rel at ed i ndi vi dual s t hat resembl e each other and are able to br eed among t hemsel ves but usual l y not wi t h other species. tetrapolar - condi t i on r ef er r i ng to sexual compat i bi l i t y of some Basidio- mycet es in whi ch two sets of f act or s are involved (such as A. a & B, b). thallus (pi . t h a l l i ) t he undi f f e r e nt i a t e d body of t he f ungus : the my- cel i al mass. veil t he partial vei l is a coveri ng t h a t ext ends from t he unopened mar - gin of t he mushroom cap to the stalk, and t hat r upt ur e s to form the annul us ; the universal veil is a tissue surroundi ng the ent i r e develop- ing mushroom, usually lost early in devel opment . Stropharia cuben- sis has bot h a partial and universal veil, but the l at t er is seldom ob- served in the absence of a magni fyi ng glass. 63 NOTES OTHER AND/OR PRESS BOOKS: HISTORY OF COCA -Mortimer S8.50 LAUGHING GAS -Shedlin, Salyer, W all e chin sky $3.50 HIGHEST QUALITY INDOOR/OUTDOOR MARIJUANA GROWER'S GUIDE -Frank & Rosenthal $3.50 COCAINE CONSUMER'S HANDBOOK -David Lee $4.95 PSILOCYBIN: MAGIC MUSHROOM GROWER'S GUIDE -Oss&Oeric S4.95 MARIJUANA LEGAL HANDBOOK -James Carroll $3.95 (available June 1976) AVAILABLE FROM YOUR LOCAL BOOKSELLER or Send Re t a i l Price plus 50^postage & handl i ng t o. AND/OR PRESS P.O. Box 2246 Ber kel ey, Ca. 94702