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Agar isolations ? * 1
    #872206 -

Whenever i receive a syringe in the mail from any of these vendors it is a particular species. But not so much a pure strain of that species? And this can only be obtained by isolating and reisolating in agar mediums until its really rhizemorphic growth? Then that is when you have the true isolation of a pure species??? Can you reisolate this pure/semi pure isolation to maybe make it tend to grow faster (i.e. take the first ropey looking strand in the mycellium and isolate over and over) Would this train the mycellium to colonize faster or is there a ceiling limit on how fast that the mycellia can colonize? Really interested in agar cultures and isolations of pure strains...


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: socratesmind]
    #872406 -

I strongly suggest you to buy the Mushroom Cultivator( Link at www.shroomery.org/books ). It will answer all and more of your questions.


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Agar isolations ? [Re: Anno]
    #872848 -

Could you maybe post the TOC or table of contents for me to see what all the book contains. I'm really interested in fungi in all aspects not jsut the magic producing ones...


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: socratesmind]
    #872918 -

Trust us, just buy it.
That and GGMM together make up a fairly complete collection of what you need to know..


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If I made affront, I apologize.
If I made affirmation, I apologize.
I merely came to listen, came to say.

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Re: Agar isolations ? [Re: socratesmind]
    #873135 -

Did you check the link I gave you at all??

One more time:

www.shroomery.org/books

At the bottom of the page......

There you will find the table of contents......


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

Edited by Anno (09/09/02 06:16 PM)

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Re: Agar isolations ? [Re: socratesmind]
    #873459 -

If you continue to reisolate the strain systematically on the same media it will get pretty damn fast, but it will also become selective and adapt the media - making its growth on other nutrients other than MEA, etc agar less viable.


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Re: Agar isolations ? [Re: Zen Peddler]
    #873486 -

So, would it be a good idea to isolate some rhizos, then grow it out on wood, and then take a chunk of colonised wood and put this on some agar again? Or would this have no effect on helping reduce substrate preferences?

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Re: Agar isolations ? [Re: Peace_Patrol]
    #873506 -

What i think bluemeanie means is that the more and more faster u stimulate it's growth its just gonna grow that much more picky when selecing a substrate other than MEA. So it's gonna be a good colonizer yes in agar and might not colonize lets say BRF or grains or stuff like that. Future experiment would be nice so we can have hard data.


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: socratesmind]
    #873513 -

Yeah, I understood that, so I was asking, would it be a good idea to avoid this single-substrate preference by sometimes growing it out on wood or some other substrate before switching back to agar.

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Re: Agar isolations ? [Re: Peace_Patrol]
    #874603 -

Well one way i was thinking that you could accomplish this to make it diverse on substrates is gradually introduce your target substrates at certain levels in the isolations and increase the substrate level with each interval.  So it will be at least use to seeing/eating that particular substrate.  This should be a nice experiemnt to try.  I'd love to do but i need to get my glovebox runnin and get some agar/petri dishes and a few other goodies :frown:


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: socratesmind]
    #875069 -

Glovebox is old school.. way old school.. if your spending money, go with a good flow hood.
have you read tmc and ggmm yet?
do so.. it will answer all these questions..


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If I made affront, I apologize.
If I made affirmation, I apologize.
I merely came to listen, came to say.

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Re: Agar isolations ? [Re: Zen Peddler]
    #875410 -

surely the problem with becoming dependent, or at least preferring a particular substrate only occurs through successive multispore innoculations?
ie if you make a multispore innoc on eg MEA, fruit the fastest growing substrain, then use the spores from this substrain in a new multispore on the same medium, fruit the fastest of these etc, then eventually, theoretically at least, you could end up with a sub-sub.....substrain that will only grow on MEA and nothing else?
If a particular substrain has the required DNA to produce the enzymes to digest a wide variety of substrates then surely no end of cloning is going to change this? Obviously if a substrain "gets used to" a particular medium it will take a while for it to start producing the relevant enzymes which were previously unneeded, eg how it takes a few days for mycelium to adapt to peroxidated agar, but I don't see how it could lose this adaptability altogether without genetic change being involved.
or am I missing something?


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hmmm........

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Re: Agar isolations ? [Re: socratesmind]
    #875558 -

the thing is there is a limited amount of time that a strain will work it doesent die but it becomes useless this is why pepole keep stock of strains in a fridge once you get it working well do this and that book has a lot of info on it


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the above is what the litel leprecans told me to say and should be regarded as such
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Re: Agar isolations ? [Re: Champion des Champignons]
    #875904 -

Stamets suggests that each transfer of an isolate should be done on new nutrient bases for agar plates otherwise the mycelia will quickly adapt to become selective. Im quoting stamets when im talking here, not from personal experience as ive noticed little difference between second generation and 12th generation on MEA


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Re: Agar isolations ? [Re: Zen Peddler]
    #880313 -

I have a question, when the agar plate has completely colonized, and it's time to examine the mycelium carefully in order to find a rhizomorphic section to transfer to the next plate. Can u just take the top off to examine w. out having to worry about contams, meaning, once it's colonized, is it pretty resistant to contams just like a colonized jar?? My friend has a plate that will be done soon and he wanted to know about this. Thanks!!!!

Vis


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"15 minutes in freezing conditions?!?! You'd become a popsicle before you made that!!!"

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Re: Agar isolations ? [Re: Visigoth]
    #885243 -

Better safe than sorry why try to contam your culture at this point just take the same precautions and be careful and remember to store your pure culture in a slant so u can always go back to it :smile: 


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: socratesmind]
    #885285 - 

code:
  
Mushroom Cultivator
TABLE OF CONTENTS


FOREWORD by Dr. Andrew Weil                                   xi
PREFACE                                                       xii
I.INTRODUCTION TO MUSHROOM CULTURE                            1
An Overview of Techniques for Mushroom Cultivation            3
Mushrooms and Mushroom Culture                                4
The Mushroom Life Cycle                                       6
II.STERILE TECHNIQUE AND AGAR CULTURE                         15
Design and Construction of a Sterile Laboratory               16
Preparation of Agar Media                                     19
Starting A Culture from Spores                                23
Taking a Spore Print                                          23
Techniques for Spore Germination                              24
Characteristics of the Mushroom Mycelium                      25
Ramifications of Multispore Culture                           25
Sectoring: Strain Selection and Development                   31
Stock Cultures: Methods For Preserving Mushroom Strains       37
III.GRAIN CULTURE                                             41
The Development of Grain Spawn                                42
Preparation of Grain Spawn                                    45
Spawn Formulas                                                46
Inoculation of Sterilized Grain from Agar Media               48
Inoculation of Sterilized Grain from Grain Masters            49
Alternative Spawn Media                                       54
Liquid Inoculation Techniques                                 55
Incubation of Spawn                                           57
IV.THE MUSHROOM GROWING ROOM                                  61
Structure and Growing Systems                                 62
Structure                                                     63
Shelves                                                       64
Trays                                                         65
Environmental Control Systems                                 66
Fresh Air                                                     66
Fans                                                          68
Air Ducting                                                   70
Filters                                                       70
Exhaust Vents                                                 72
Heating                                                       73
Cooling                                                       73
Humidification                                                74
Thermostats and Humidistats                                   74
Lighting                                                      74
Environmental Monitoring Equipment                            76
V.COMPOST PREPARATION                                         77
Phase I Composting                                            78
Basic Raw Materials                                           78
Supplements                                                   79
Formulas                                                      81
Ammonia                                                       82
Carbon:Nitrogen Ratio                                         83
WaterAir                                                      83
Pre-Wetting                                                   84
Building the Pile                                             85
Turning                                                       87
Temperature                                                   88
Long Composting                                               89
Short Composting                                              90
Synthetic Compost Procedure                                   91
Composting Tools                                              92
Characteristics of the Compost at Filling                     93
Supplementation at Filling                                    95
Phase II Composting                                           96
Basic Air Requirements                                        97
Phase II Room Design                                          98
Filling Procedures                                            98
Depth of Fill                                                 99
Phase II Procedures: Trays or Shelves                         100
Phase II in Bulk                                              101
Bulk Room Design Features                                     102
Bulk Room Filling Procedures                                  104
Bulk Room Phase II Program                                    104
Testing for Ammonia                                           104
Aspect of the Finished Compost                                105
Alternative Composts and Composting Procedures                106
Sugar Cane Bagasse Compost                                    106
The Five Day Express Composting Method                        106
VI.NON-COMPOSTED SUBSTRATES                                   109
Natural Culture                                               110
Wood Based Substrates                                         114
Straw                                                         117
VII.SPAWNING AND SPAWN RUNNING IN BULK SUBSTRATES             121
Moisture Content                                              122
Substrate Temperature                                         122
Dry Weight of Substrate                                       122
Duration of Spawn Run                                         124
Spawning Methods                                              124
Environmental Conditions                                      125
Super Spawning                                                126
Supplementation at Spawning                                   126
Supplementation at Casing                                     126
VIII.THE CASING LAYER                                         127
Function                                                      128
Properties                                                    129
Materials                                                     130
Formulas and Preparation                                      132
Application                                                   133
Casing Colonization                                           135
Casing Moisture and Mycelial Appearance                       137
IX.STRATEGIES FOR MUSHROOM FORMATION (PINHEAD INITIATION)     139
Basic Pinning Strategy                                        140
Primordia Formation Procedures                                141
The Relationship Between Primordia Formation and Yield        146
The Influence of Light on Pinhead Initiation                  147
X.ENVIRONMENTAL FACTORS: SUSTAINING THE MUSHROOM CROP         149
Temperature                                                   150
Flushing Pattern                                              150
Air Movement                                                  152
Watering                                                      154
Harvesting                                                    155
Preserving Mushrooms                                          156
XI.GROWING PARAMETERS FOR VARIOUS MUSHROOM SPECIES            159
Agaricusbitorquis                                             161
Agaricus brunnescens                                          164
Coprinus comatus                                              168
Flammulina velutipes                                          172
Lentinus edodes                                               176
Lepista nuda                                                  180
Panaeolus cyanescens                                          183
Panaeolussubbalteatus .                                       186
Pleurotus ostrealus (Type Variety)                            189
Pleurotus ostreatus (Florida Variety)                         193
Psilocybe cubensis                                            196
Psilocybe cyanescens                                          200
Psilocybe mexicana                                            204
Psilocybe tampanensis                                         207
Stropharia rugoso-annulata                                    210
Volvariella volvacea                                          214
XII.CULTIVATION PROBLEMS AND THEIR SOLUTIONS:
TROUBLE SHOOTING GUIDE                                        217
Sterile Technique                                             219
Agar Culture                                                  219
Grain Culture                                                 220
Compost Preparation                                           223
Phase I                                                       223
Phase II                                                      224
Spawn Running                                                 226
Case Running                                                  227
Mushroom Formation and Development                            229
Pinhead Initiation                                            229
Cropping                                                      231
XIII.THE CONTAMINANTS OF MUSHROOM CULTURE:
IDENTIFICATION AND CONTROL                                    233
A Key to the Common Contaminants of Mushroom Culture          238
Virus (Die-Back Disease)                                      244
Actinomyces (Firefang)                                        246
Bacillus (Wet Spot)                                           248
Pseudomonas (Bacterial Blotch & Pit)                          252
Streptomyces {Firefang)                                       255
Alternaria (Black Mold)                                       257
Aspergillus (Green Mold)                                      259
Botrytis (Brown Mold)                                         262
Chaetomium (Olive Green Mold)                                 264
Chrysosporium (Yellow Mold)                                   266
Cladosporium (Dark Green Mold)                                268
Coprinus (Inky Cap)                                           270
Cryptococcus (Cream Colored Yeast)                            273
Dactylium (Cobweb Mold)                                       275
Doratormyces (Black Whisker Mold)                             277
Epicoccum (Yellow Mold).                                      279
Fusarium (Pink Mold)                                          281
Geotrichum (Lipstick Mold)                                    284
Humicola (Gray Mold)                                          286
Monilia (White Flour Mold)                                    288
Mucor (Black Pin Mold)                                        290
Mycelia Sterilia (White Mold)                                 292
Mycogone (Wet Bubble)                                         294
INeurospora (Pink Mold                                        296
Papulospora (Brown Plaster Mold)                              298
Penicillium (Bluish Green Mold)                               300
Rhizopus (Black Pin Mold)                                     302
Scopulariopsis (White Plaster Mold)                           304
Sepedonium (White or Yellow Mold)                             306
Torula (Black Yeast)                                          308
Trichoderma (Forest Green Mold)                               310
Trichothecium (Pink Mold)                                     313
Verticillium (Dry Bubble)                                     315
XIV.THE PESTS OF MUSHROOM CULTURE                             319
Mushroom Flies                                                320
Fly Control Measures                                          320
Sciarid Fly                                                   321
Phorid Fly                                                    323
Cecid Fly                                                     325
Mites                                                         328
INematodes (Eelworrns)                                        331
XV.MUSHROOM GENETICS                                          333
Reproductive Strategies                                       336
Implications for Culture Work                                 338
APPENDICES                                                    343
I.Medicinal Properties of Mushrooms                           345
II.Laminar Flow Systems                                       347
III.The Effect of Bacteria and Other
Microorganisms on Fruiting                                    253
IV.The Use of Mushroom Extracts to Induce Fruiting            357
V.Data Collection and Environmental Monitoring Records        359
VI.Analyses of Basic Materials Used in Substrate Preparation  369
VII.Resources For Mushroom Growing Equipment and Supplies     384
VIII.English to Metric Conversion Tables                      386
GLOSSARY                                                      389
BIBLIOGRAPHY                                                  397
INDEX                                                         409
PHOTOGRAPHY AND ILLUSTRATION CREDITS                          414
ACKNOWLEDGEMENTS.                                             415


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Agar isolations ? [Re: Zen Peddler]
    #885983 -

In reply to:
If you continue to reisolate the strain systematically on the same media it will get pretty damn fast, but it will also become selective and adapt the media - making its growth on other nutrients other than MEA, etc agar less viable.



Well, could you make some brown rice flour agar and keep isolating on it, then spawn it to some whole brown rice jars? Won't it be totaly suited for that substrate? Thus growing faster and better on it.


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Everything posted above was out of fun, none should be taken serious. I am currently under the influence so take that into consideration.

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Re: Agar isolations ? [Re: Anno]
    #886489 -

Thanks a mucho Anno i think i'm going to B&N to get my book special ordered tomm :smile:


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: socratesmind]
    #886903 -

>i think i'm going to B&N

Is the book cheaper there?
I looked and the book is not available there
http://search.barnesandnoble.com/booksearch/isbnInquiry.asp?userid=0DDKSXPISD&isbn=0961079800

You can order it at Amazon:

http://www.amazon.com/exec/obidos/ASIN/0961079800/minmed-20/ 


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Agar isolations ? [Re: socratesmind]
    #886947 -

You should buy your books through the Shroomery!

Help support the site.  :smile:

 


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Laterz, Road

Who the hell you callin crazy?
You wouldn't know what crazy was if Charles Manson was eating froot loops on your front porch!

Brainiac said:
PM the names with on there names, that means they have mushrooms for sale.


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Re: Agar isolations ? [Re: Roadkill]
    #887431 -

Well B&N can get any book as long as you have the ISBN and I'd love to support shroomery but I get a much needed discount on any book thru B&N. But thanks again guys/girls.


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: Visigoth]
    #889963 -

sure it shouldnt contaminate but it will no longer be a pure culture in that it will be riddled with contaminant spores and these will over-run any substrate that you use the wedge to innoculate.
PF is a good example of a strain that has been selectively grown on brown-rice flour and cakes - it performs poorly on other media


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Re: Agar isolations ? [Re: socratesmind]
    #889974 -

>Well B&N can get any book as long as you have the ISBN

Didn?t you read my post? It?s OUT OF STOCK there...
http://search.barnesandnoble.com/booksearch/isbnInquiry.asp?userid=0DDKSXPISD&isbn=0961079800


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: Agar isolations ? [Re: Anno]
    #890836 -

Well these 4 used copies? http://search.barnesandnoble.com/OopBooks/OopResults.asp?title=The+Mushroom+Cultivator%3A+A+Practical+Guide+to+Growing+Mushrooms+at+Home&author=Paul+Stamets&userid=5114PUBNMQ


--------------------
Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.

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Re: Agar isolations ? [Re: socratesmind]
    #891279 -

Hahaha, did you see the price of those used copies and compared it with the amazon price?
http://search.barnesandnoble.com/OopBooks/oopResultsTitle.asp?userid=1A3WYTRKC9&author=Paul+Stamets&title=The+Mushroom+Cultivator%3A+A+Practical+Guide+to+Growing+Mushrooms+at+Home&rstart=1&WID=1219248


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Shop: Myyco.com Isolated Cubensis Liquid Culture For Sale   MagicBag.co All-In-One Bags That Don't Suck   North Spore Bulk Substrate   Bridgetown Botanicals CBD Concentrates   Unfolding Nature Unfolding Nature: Being in the Implicate Order   Mushroom-Hut Liquid Cultures   Original Sensible Seeds Bulk Cannabis Seeds   Left Coast Kratom Buy Kratom Capsules   PhytoExtractum Kratom Powder for Sale

Mushrooms, Mycology and Psychedelics > Advanced Mycology
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