My Cultivation technique BRF + honey + creating your own spore syringes,Building your Terrarium,Inoculating your jars :)

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EskaTaRi  bLu   11/02/10

This is a walkthrough of my 1st attempt at growing psyilocbe cubensis,specifically the B+ strain via the BRF technique involving the use of jars and spore syringes, Written down whilst sterilizing and inoculating jars, All knowledge was written from information i studied and memorised over a period of several months where i became deeply intrigued by the world of mycology. The main reasons that fuelled this obsession was my love for physcadelic substances and how they react on peoples minds, Make them feel, All of which are triggered by various chemical compounds in our brains and bodies which are even more interesting as you try and understand what they are, and how they work.

Now lets not get sidetracked, The purpose of this document is like a draft for the mycologist brains that take the time to read this, and i wish my mistakes to be corrected :)  Although i hope all goes well and my 4 inoculated jars remain contaminant free and yield a large, potent crop of Cubies and Azursecens.

(Ps note I’m retyping my notes exactly so measurements and amounts  used are rough if not even mentioned at all, I just used equal amounts off all materials needed for the substrate, vermiculite, Plain white flour, Wholemeal long brown rice, And 4 teaspoons of honey.)

(**) Sterilize cup, open spore print and scrape either a small amount of spores, or the whole lot depending on desired harvest size, Number of jars to inoculate etc, Use a clean sterile razor blade for scraping  ,( I’m using Wilkinson’s swords razors, they come in a tiny box, individual and sterile, Also very sharp! 25 a pack, $4.50 Coles)  Do not expose spore print to the surrounding air for too long, Make sure that you also sterilize the water in the microwave until it reaches boiling point, Boil for 4 mins. Remember to allow water to cool if its too hot it will burn and kill your spores, when it cools to room temperature it should be fine, around 25c should be fine, Now back  to the scraping of the spores from your print, For best approach sterilise water and cup before opening . Use a 5ml syringe to measure how much you are going to need,1 print can make 10 five ml spore syringes I’ve been told ,S0 maximum amount of water needed is 25ml., 1 spore syringe can inoculate 2-3 jars ,So using 10ml water with 1/4  of a spore print should be fine and will be sufficient to inoculate 6 jars, of course u could use the whole print for more jars(like I have done), Roughly 10 syringes derived from 1 decent spore yielding print, can inoculate 22 jars, Figuratively speaking?

(2) Mix BRF (brown rice flour) +LGR (long grain rice) in bowl. Put vermiculite in a bowl and soak in water (Necessary for water  source for developing mycelium) Mix the BRF and LGR ,Mix well also you can add a few spoonfuls  of honey  for extra nutrients , mycelium apparently loves glucose. now begin Packing the jars well with substrate (BRF+ LGR + optional honey, or pure glucose gel?) fill the jar halfway up with water and pack the jar almost to the top, leaving about half inch  or inch and place DRY vermiculate in and pack relatively well, but not too tight,this allows gas to exchange out of the substrate, Now  if the lids have a plastic seal,  stick  new sterilized aluminium foil under your lid, and mould it to the shape when you are preparing to reseal the jar for sterilization, Next using a half inch sharp nail(,Preferably pre soaked in bleached and washed in boiling water,) Were going to punch 3 holes  in a triangle fashion. Tear off 2 descent strips of aluminium foil , place over lid punch 3 holes into the 1st foil strip , Now cover  the 1st strip with  another foil strip , securing tightly, To prevent bacteria from entering via our inoculation holes, I used electrical tape and tightly taped around the lid for added peace of mind,

Now  Fill a saucepan with water until its equivalent with the jars water level, make sure you put a cloth under the jars to absorb some of the heat ( this stops the jars from cracking )  boil for 2, 2 1/2 hours at boiling temperature, and monitor drops in water levels and refill as needed, Once done remove the jars and  leave to cool, For best results  let jars for 24 hours  , this way we can see if any bacteria  has survived sterilization , look for mould spots , anything irregular  will show up ,  We can sterilize again  for the same time as before ,  And once again allowing them to cool and leaving them for 24 hours, Checking them again for bacterial growth , if its nice and clean IE no mould spots or other types of strange spots, We can begin inoculation . BEWARE THOUGH jars must be completely free of any competing bacteria or mould, mushrooms will still grow if infected with bacteria spores or mould, but will make you very sick and sometimes fatal. Depending on the toxins in the bacteria, etc etc, Sterilizing twice over 48 hours is the best approach, (but once can also prove sufficient enough) to make sure its sterilized and viable to use. Some endospores and bacteria, moulds etc cant be killed boiling temperatures, So  expect to lose at least 2 jars for every 6 you do, It may not happen, If you remember to keep everything sterile, its a simple process, But fumble any part of the experiment where it be preparing substrate to inoculating jars to preparing spore syringes from spore prints, And it can ruin jars for good, Even whole batches if the spore solution used for inoculation is in any way exposed to contaminents,dirty hands, airborne particles.bacteria,spores etc, the list goes on. But i've heard on a few mycology documentaries even professional mycologists expect to lose 2 jars, or substrate bags per 6 substrate bags/Jars.

(3) Now its time to inoculate our sterilized jars, Grab a clean new sterile 5ml syringe (  a 5ml Sterile syringe, easily obtained from any pharmacy for 30 cents, And I used sterile needle tips with gauges ranging from 18G to 21 Gauge) I used a few so obtaining 4-5 spare of 21Gauge,20gauge,and 18gauge is worth the few dollars they cost, Some may clog straight up ,So best response I found was throwing the clogged one aside in a cup of bleach, attaching a new sterile tip, flaming the needle tip and 90%of its metal tip with  lighter ( I only used a regular Bic lighter, Professionals suggested to use a jet lighter, they don’t produce carbons so i’ve been told) And returning back to the inoculation point where the tip gets clogged, Now just utilise the small tunnels that were carved previously taking care to go slow and tediously to avoid clogging the finer gauge tips (21 -20gauge) To make things abit easier I found that if you get a clogged tip (better if its a larger gauge,EG 18G,19G,20G) just sterilize it in a cup of bleach for 10 mins,steralize hands in bleach solution ( wearing gloves!) and then wash hands and needle tip both under very hot water , u can use it to create the pathways at the inoculation holes, so it minimizes the chance of the spore syringe getting clogged with substrate, And then risking contamination of the spore solution by increasing exposure to airborne particles ,and in general the surrounding environment, when you would need to change needle tips, So this tip is indeed worth the extra time to sterilize a needle tip, and maintain its sterilization while carving tunnels for the spore solution to travel through, The key to success is never forgetting to flame the needle tip each time its going to be placed through those inoculation points, and again when its taken out  to change angle or begin inoculating another entry point. Squirt spore solution via the routes made until it runs down the sides of the glass, if using 5ml solution about 04ml interval squirts, 3 times, each of the 3 inoculation points, kind of  like /|  at each inoculation point ,And my technique I used here, was one entry   one a angle to the left, then another entry  / at the reverse angle to the right, then another just straight down, to maximise the surface exposed to the spore solution.

Once mycelium starts to develop, it will resemble a miniscule amount of milky residue in scattered places through out the substrate, As time goes on it will grow into large often round pieces of white mycelium flesh, Eventually it will colonise the whole jar including the bottom which is normally last. This can take 1 week to 3 weeks even longer, How well substrate was mixed, ratio's used, temperature during incubation, they all affect the duration of time this will take, Poorly constructed substrates may never fully colonise. Be on the lookout for any unusual spots, marks etc. The only safe marks are bluish marks, bruises or spots, or rarely sometimes a tinge of yellow, from lack of nutrients or something along those lines. The bluish marks bruises spots etc anything blue is a reaction from contact with the psilocybin and a foregein object, like our hands amino acids will cause this reaction. Discard any jars which contain anything other than white mycelium. Or slightly tinged yellow. Research appropriately before assuming its safe.

When ready for transfer into a terrarium once fully colonised and established turn upside, Tap bottom and it should slide, right out I’ve read that mycelium shrinks after its established on the BRF cake. Transfer immediately into a terrarium with a perlite bottom and regulated humidity at levels of between 88% RLH and 89% RLH, For successful fruiting water 3 times daily avoiding spraying the mycelium directly (allow 1- 6 hours of light per day) they can survive on 1 hour per day, Fruiting will take 3 - 7 weeks, or it will continue until substrate nutrients are depleted.

Soaking perlite in water is a good easy way to control humidity, once sealed it is sucked back out by surrounding air and then sticks to the glass creating artificial humidity .

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            Building a Terrarium.

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Items needed:

* Plastic greenhouse used to grow seedlings, $10.00 bunnings...Relatively cheap, But also really thin and crap plastic, It probably would work But a better alternative is a large plastic storage container roughly 1 meter long and 40-50cm high and 40 cm’s wide across the sides, Size will vary depending upon how many jars you have for transfer, How much the specific strain yields, Size of mushrooms etc,But those measurements should provide a basic estimation for 4-6 jars of Most P.cubensis strains

*HYGROMETER $17.00 (ebay.com.au) that reads between 30-40% Relative humidity and 98% Relative Humidity+ accurately analyses surrounding temperature, This device is critical to the success of your crop fruiting, As the mycelium feeds of the moisture in humidity, this is the lifeline off the mycelium, Its so critical to keep the humidity at a constant 95% or above because the mycelium cannot be watered directly, as the water droplets would damage the fungus I have been led to believe, Tho I have not seen It with my own eyes yet, Time will tell J

*Bag of PERLITE growing medium, $10.00 Bunning’s

*Water spray bottle, with fine mist setting.

*1m X 1m Clear Perspex sheet (For terrariums mist guard) not sure on price.

* 1 clean large cardboard box

* OPTIONAL Steam Humidifier.

 (B) Once a suitable object is Ascertained for our terrarium, Take the perlite and soak it in water until the perlite absorbs the water that was poured onto it, A good way to tell when the perlite has absorbed its maximum amount of water, Is that it will start to pool in the bottom of the bowl, Along with all the water which could not be absorbed. Now after sufficient water has been absorbed take the perlite and line the bottom of your chosen terrarium Box, Im going to use a large sterile storage container from bunnings, they are roughly the size of a decent esky that would hold around a carton of stubbier beers (30 beers/50L) so that should give you a rough mental image of the containers size.. I forget to mention earlier that the lining of your terrariums bottom with perlite should be done 24-48 hours before the transfer of our BRF cakes from our Inoculated jar, This way our terrarium has had 1 or 2 days to regulate its humidity to the desired level of 98%, And we can also use this time to monitor humidity levels and see if we can keep it at a constant RLH above 88% RLH.Using an accurate hygrometer is crucial. And also to make sure the temperature is a nice 28c for our P cubensis B+’s. Following these tips will ensure  our transplanted mycelium is transferred directly into a perfect stable environment, Of minimum 88% RLH and 28C this will promote optimum growth times and optimum fruit yields and quantity of fruits, Potency as well I presume.

Once our mycelium has fully colonised the BRF substrate cake completely, Back, sides, bottom, It is generally told to be Quiet resilient to contamination from bacteria, Mould endospores ,Floating particles, and micro organisms, because its already established and fully covering the substrate cake, vitally protecting all the vulnerable substrate nutrients inside the BRF cake. Keep in mind that the Mycelium on the BRF cake cannot be sprayed directly with water no matter how fine the spray mist is. To overcome this obstacle, Acquire A 1m X 1m piece of clear Perspex sheet  and some very strong glue, perhaps an epoxy or lock tight, After we transfer our mycelium into the terrarium , just about one ½ half inch below the terrariums lid cut and shape the Perspex to fit there, and glue into place, Making sure its angled on a 25 degree tilt to block water spray mist from touching the mycelium, but allowing to touch the sides of the terrarium and allowing the water droplets to run of into the perlite below to be absorbed, , securing with a strong teck screw of needed for stability issues..

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Hygrometer must be constantly monitored on a daily basis, to make sure the humidity levels stay above 88% RLH, Monitoring Humidity levels basically at this stage are one of only a few caretaking daily duties needed to be undertaken without any mistakes. The only other duty is  spraying into the terrarium with a water spray bottle 3 times  day, with it set on a very fine mist, After performing this duty,  Take the lid and use it to fan above the terrarium to fan out lingering C02 produced constantly by the fruiting mycelium, If its allowed to pool up in the terrarium it will cause yield problems, yield times, Hell it can even stop the mycelium fruiting at all, Since Fruiting is triggered by decreased  C02 amounts almost instantly after its taken from the sealed inoculation jars, Where for so long it needed the opposite, Lots of C02 during vegative state.

 Decreased C02 tells the mycelium that the environment has changed and it must start to bear fruit. Fruiting should start to appear within 1 week of transfer from jars, but has been known to show the preliminary stages of fruiting mushroom heads while still contained in inoculation jars, though I’ve learned this should not always be expected. Fruiting will continue until all off the nutrients contained within the BRF substrate cake have been depleted, this included the long grain rice, flour,  and Honey if you added it.

In my batch I used honey as a nutritional supplement , supposedly it supplies the mycelium with higher levels of glucose ( Sugar ) and is said to increase potency, Total mushroom yield And size all in all, I mixed about 3-4 teaspoons of honey into my substrate before sterilization and I’m eagerly waiting to see if it worked or if it fucked it, I think I sterilized for too long ( 2 and half hours) because I can see on the sides where the honey has caramelized and turned a rich brown colour generally predominant through the bottoms of all my 4 jars. Another worry I had was that if I indeed did sterilize for too long it may have allowed the water contained in the vermiculite within the substrate itself to evaporate, leaving the inoculation jars without a water source for the mycelium to feed of. However knowing the jars were well sealed during the sterilization/Boiling stage, im confident not much if any water was able to leave the jars. I will just have to wait it out to know the truth

PRICELIST FOR STERILIZATION & INNOCCULATION PROCESS & TERRARIUM CONSTRUCTION + MISC ITEMS

• Five 5ml sterile syringes from local pharmacy : 30c each, 5 x 0.30 = $1.40 AUD
• Vermiculite: $6.00 from Bunning’s
• Home brand Aluminium foil $1.50 - $1.90 COLES
• Sterile rubber cleaning gloves $3.00 pack of 3 pairs @ Coles
• Home brand Stir fry jars, medium size approx 540grams No tapered inwards jars or retarded jar designs to make transferring easier on yourself, These jars are pretty average but will do the job. I bough 4, they cost $1.85 each
• 4 types of these needle tip gauge sizes, 21 gauge, 20 gauge 18 gauge , they are cheap and its definitely worth having a few of each size laying around handy.
• 2L of home brand bleach :   $ 2.00
• Wilkinson’s sword double sides razor blades  $4.00 but pocket sized for five finger discount, + you get over 20 blades individually wrapped to at least each comes clean, sterile and ready to scrap spores etc.
• Electrical tape
• Sterilized shot glass for holding spore solution when make spore syringes.
• Distilled water for suspending of spores in water and for substrate mix
• Honey.
• AJAX heavy duty surface disinfectant and anti bacterial spray (ITS PINK) unsure of price
• Large mixing bowel
• Decent sized saucepan for boiling jars in, At least over 8cm tall sides, Enough to allow jars to sit in there on top of a tea towel(This helps to stop jars getting to hot and cracking), but  only to the middle point of the jars. Where our water line stopped when we filled up our jars halfway with water while introducing our substrate mix to the jars before sterilization, Sounds easy enough, don’t forget!
• LARGE plastic storage contained roughly 1m long 50 cm tall, 40cm wide. Around 50 litres, the average camping esky
• 1 water sprayer with fine mist setting
• 1m X 1m clear Perspex sheet
• Decent strength glue
• Bag of perlite
• Hydrometer
• OPTIONAL humidifier $40.00
• _____________________________________________-

                 Generally for batches of 4-6 jars the cost should be no higher than $20.00, unless you buy brand name crap.

Data collected from daily observation of inoculation entry points on each jar, As well as area’s which seem to be developing milky spots and blobs of mycelium at its early stage of development.

Wednesday 10^th February 2010

I inoculated my jars on Sunday 7^th February 2010 between the hours of 12:30am and 02:30am

*I inoculated 4 jars *** 3 JARS WITH PSILOCYBE CUBENSIS “B+” STRAIN

*** 1 jar with PSILOCYBE AZURESCEND “HAMMOND STRAIN”

USING 1/3rd of its print for 1 single jar. Using 1/3 of its print I made a 5ml solution carefully following sterilization techniques and injected it into all 3 inoculation points of 1 of our jars,  Carefully avoiding contamination (Remembering to Flame 90% of needle tip till red and then cooling under cold sterile water and doing it each time it comes and goes from entering the shot glass and the sterile holes in the jar  lid ) I used a further sterile 5ml of water and sprayed it into the shott glass I used to hold my spore solution as well is mixing the spores thoroughly throughout the water, This enabled me to suck up every stray spore that was scrapped into the shot glass or escaped the first solution I sucked up, Be observant spores my have a hard time completely leaving our spore syringes ,typically at the front were the solution is pushed down into the needle is where I saw it collection, Sterilize tip, suck up sterile water and shake syringe vigorously until the tiny spores are freed, If not, Don’t stress, Many times more spores just entered the inoculation jars