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Hey guys, I've been having a rough time learning to use spawn bags and I could use some help. I'll describe my procedure, and what happens, and you guys tell me where I'm going wrong.
I start with 3 pounds of WBS, and my first step is to add enough water to cover it with about 3 inches of space. I use a colander to skim off any floating seeds, then work with my hands to move around all the seeds in the bowl of water, skimming as I go. The next step is to strain the seeds, add more water, work with hands, rinse, repeat, until the water no longer clouds up. I then add hot water until the seed is covered with about 2-3 inches of clearance, and then let it sit for 24 hours. During this time, I'm told than the heat resistant sour patch spores that may be on them will germinate within 24 hours, allowing me to kill them off.
The next day, when I see signs of bubbles and a phenolic 'hammy' odor characteristic with the mold type, I redrain the seeds, give them all a good rinsing, then with a small colander move bit by bit, using my hands to rub the seed under the water until they no longer feel slimy. When I'm finished, the seeds are about 2 shades lighter than how I started. This goes into a pot with hot water and allowed to raise to 200f, where I keep the temp constant for 15 minutes and allow the seeds to swell up, but not explode. Once the seeds reach a theshold I've come to identify as 'almost gonna burst' I strain the seeds and add them to a bowl of icewater. This sits for 5 minutes of constant stirring.
I allow the seeds to strain for about half an hour, then add all three pounds to my spawn bag, filling halfway to the filter patch. I then use a foodsaver to heat seal the bag shut, wrap it around itself, leaving the filter facing 'up', and weigh it down with a plate. This does into the PC on top of metal jar lids with a little water, digitally controlled to 15 PSI for 75 minutes. I take out the bag once complete and the pressure's dropped and hang it up to let the bag cool.
Once cool, I prepare a syringe in my glovebox by wrapping the syringe in foil and autoclaving it for 15 minutes. I then take the time to sterilize my box and the room I'm working in, and take a quick shower. I put on my gloves and transfer and foil-wrapped syringe into the box. From there, I remove the plastic tip and flame sterilize the needle, take the filter disc off my lc I'm using(I have burma and ecuador. I sample from whichever one has the most material in it) and while it's waiting, I keep a square of alcohol swab over the injection hole. Immediately after flame sterilizing the needle, it's wiped with an alcohol swap, and poked through the alcohol swap on the lc jar and into the medium. For each bag I use about 5ccs of LC. I remove the needle, cap it, sterilize the surface of the filter disc and replace it. From there this comes out of the glove box and I'm ready to inject.
This step is the easy part. I flame sterilise the needle, wipe it with with a swab, wipe down the bag's self-healing injector site with a swab, and inject. From there it goes into my martha, where I keep a temp of 80-82 degrees fulltime to let it sit and think about what it did.
I have done this at least 6 times. Each time, I either start to see a few signs of colonization, or none at all, and then it goes wrong. I start to smell a sour garbage-like or phenolic ham character. Any part in the bag where there's moisture I see it starting to increase and turn and ugly brown color. Eventually the bag stops colonizing, doesn't colonize at all, or the colonizations ceases to be visible to the naked eye.
I've been making beer and Sake for 5 years, and I consider my sanitation techniques to not be terrible, but this isn't good. What am I doing wrong here that my bags are consistently contaminated by the exact same thing? I'm tempted to revert to the simpler 'BRF' technique, but feeling like maybe there's some step that I 'just dont get'.
Texturewise, my seed flows dryly when strained. Are both of my LCs contaminated possibly? I have 6ccs of ecuador still in the syringe. Should I try a new LC batch using silicone? What kind of solicone should I use? With consistent negative results I'm suspecting that those LCs may be my culprit(sigh). It's very disappointing to be putting 6 weeks into this so far and not seeing any positive results. For every injectionprep I resterilize my needles and change to a fresh pair of gloves I wipe down with an alcohol swap. When 'eyeballing' something, I keep it elevated above my nose, so I do not breathe on it.
first off i would skip the whole ice water part you want your grains hot so they can steam themselves dry
second 5cc's for 3 pounds of grain is really low i would do 10-15 and if youre using LC mix the bag up after you inoculate this will spread the mycelium around giving it more inoculation points
third the martha you dont need to put then in there just a shelf or counter top will work while colonizing
how do you make your lids for your LC? a pic would be great here
Best Thread Ever CapZilla said: not sure what GE and FAE are but i should probably get some. Citric said: Your signature is wrong on colonization temps! GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef
It's a 1/2 pint jar with a single hole in the lid and no silicone. This is sealed off from contaminants using a filter disc I got from sporeworks. I make my lid holes using a drill bit. 10-15ccs for a large bag? Oh my!
I did variants, also, on each of my bags after each successive failure.
The first batch, uncolored, was done in jars. In the first try I added cold water to the seed and rinsed it, skimmed off the floaties and let it soak in the fridge for 12 hours. I did a quick rinse and added to the jars. PC'd, allowed to cool and added a syringe a friend gave me. These never germinated and I'm writing it off as dead spores.
The second batch I had a bag. I did the same thing and I got some inoculation, but when they started to smell sour I asked around and people said 'wait and see'. That one's on week 8 and still looking prettymuch like a lost cause.
Next batch I did 3 bags using the same method. After a week they too started smelling sour. I pc'd them to kill off anything inside and tossed em in the composter.
Batch after that I was worried I wasn't using enough moisture and the grains were drying out, so this batch(colored red) I added a cup of white sticky rice. This made some firm cakes in the bag. I inoculated. After a week the firm cakes turned into red pudding that smelled like pureed garbage. They too were discarded. These I didn't skim off the floating seed, and blame contamination on those, as the air pockets could house bacteria that didn't get killed off in the PC.
After that I did a batch I dyed green. As a variant I more carefuly washed and drained, but did not allow to soak. I simply brought a pot of water up to boiling, added the seed and removed from the heat to allow to soak for 20 minutes. I strained it in how water, washed it until the liquid was clear, bagged, sealed, PC'd and innoced. It's been a week later since that point and I was seeing brown moist spots like I'd seen in previous batches. On your note on shaking the bag, I untaped it from where it was rolled up and carefully sifted through it just now and found that it's growing just peachy and is 30% coloinized. The bag's filter smells mushroomy, and not like phenolic garbage!
The batch after that I dyed yellow using the method described in the OP. All the bags smell contaminated and after 5 days I see no signs of growth.
I think I used the burma strain on the green bag and ecuador on all the others. If it's the LC, I'd say that the ecuador's contaminated. Is there a sort of mini controlled experiment I can conduct without having to prep a full bag, though? Maybe I'll do four jars with a small volume of grain in each one, with one inoculated with burma LC, one with ecuador LC, one with straight ecuador spores, and one untreated, as a control to test my sterilization technique. Does this sound good?
yea 10-15 you gotta think 3 pounds is a lot of grain i usually use 2-3 per quart jar of grains and have full colonization in about 10 days
since you have filter disks try a lid like this instead of the tyvek just silicone a small circle of filter disk there that way you dont inject thru your filter the hole filled with silicone is 1/8 of an inch the hole covered by tyvek is 1/4 inch
Best Thread Ever CapZilla said: not sure what GE and FAE are but i should probably get some. Citric said: Your signature is wrong on colonization temps! GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef
i have started making new lids using rubber injection ports and a synthetic filter disk altho the tyvek worked well too i just like the look of these lids better
Best Thread Ever CapZilla said: not sure what GE and FAE are but i should probably get some. Citric said: Your signature is wrong on colonization temps! GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef
Thanks for the info, I'll go and get some stuff to do the silicone injection site today.
Also I'm going to do an experiment. Eight halfpint wide-mouthed jars. Instead of birdseed I'll use whole rye seed. I'll do them in a batch to prep, for washing, soaking, simmering and PCing in jars. Then in two I will innoc with ecuador LC, two with burma LC, two with straight ecuador spores, and two with nothing at all, to act as my control. I'll wait and take a picture every few days. I will split up the batches to have half sitting in an 80F chamber to spawn, and half sitting on a shelf at ambient temps. This will help me isolate my problem to see whether it's my LC or my sterilization techniques. and also for the sake of saving energy, if the temperature boost gives a benefit(if any) to spawning or encourages accelerated contamination(out of the hopes that if I can't control the contam, if I can outgrow it before it gets a foothold). I will start soaking and will document each step with photos.
For 1/2 a cup of grain per jar, how much innoc should I use? 1cc?
sounds like you got a good plan there i fill my quart jars 3/4 the way full with spores i use 1cc and shoot where the grain and the glass meet do this in 3 spots and dont shake the jars this is so the spores can germinate and mate then spread
with LC i use 2-3 then shake to spread the live myc around once jars are about 30% colonized give them a good shaking then no more till youre ready to use them
Best Thread Ever CapZilla said: not sure what GE and FAE are but i should probably get some. Citric said: Your signature is wrong on colonization temps! GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef
Started the experiment 2 days ago. Rinsed the grain until there was nothing but clear runoff and soaked for 24 hours. Water turned orangish and smelled like the typical surface germination of bacteria that had an off smell. Rinsed until clear once more, and heated to 200 degrees and kept for 15 minutes until the seeds swelled up. Strained the seed and let it steam dry itself for 30 minutes(I understand what you said about the heat drying the grains. Thanks for that!). Filled up 10 jars(measured for 8, final volume of grain filled 10, so I had a surplus) and after PCing at 15psi for 75 minutes, let them cool overnight. Yesterday I prepared 2 needles with different LCs and knocked up 6 jars, 2 of them with the questionable ecuador LC and 4 with the burma LC. Hit 2 more jars with straight spores from an ecuador needle. The other 2 I left untreated for the sake of acting as a control. I split the jars in half, putting one half in my 82 degree martha and the other half on a high shelf in my room. This is to test to see if any of the LCs or spores are precontaminated, or if my sanitation techniques simply aren't working, and also to see if the theory that ambient temp is adequate for colonization. It's only been 24 hours, so I'm not seeing any signs. I'll update with any status changes.
As an update on the one 'good' bag that seemed to have healthy myc growth, the myc has stopped colonizing, and the bag is starting to take on a 'hot, sour' smell, which I assume is sour patch, as the amount of the moisture in the bag is apparently increasing with the breakdown of starches. Sad. This is all terribly discouraging. If this experiment doesn't pan out my next step would be to try a different method, i.e. BRF. From all my reading it seems a massive step backwards for efficiency, but if it works, it works. 6 weeks and no positive outcome is extremely discouraging!
To describe my sanitation, I rely on 3 elements. First one being Oust, which I spray in the room I work in thoroughly and let settle before using. The second is Sanstar, which is a concentrate of blended acids that act as an antibacterial, antifungal insecticide that is considered 'food grade'. I've used it for 3 years to sanitize beer and sake brewing materials and have never had any contaminations. This I use in a spray bottle to spray inside of my glovebox, as well as the gloves I use. The third and final is rubbing alcohol, which I have in swabs that I use to wipe down my syringes just before use, and to also rub all over my gloves, as well. When I do my work I wrap my needles in foil and pressurecook them at 15psi for 15 minutes. I then shower and dress in fresh, clean clothing with a short sleeved shirt. I wash my hands with a mixture of alcohol and antibacterial soap up to the elbows, and make sure to get under my fingernails. I rinse, and put the gloves on without drying. With my gloves on I retrieve my syringes and put them into the glovebox. This is followed by a second change of gloves which are also sterilized. For each injection, I prep the needle by flaming the tip from base to top until there is no fluid inside and the tip is glowing. I cool the tip with an alcohol swab. On my LC jars, I open the filter disk and slide a swab underneath to cover the hole in the jar. When I get my LC out, I inject the needle through the swab, pull out the measure I need and after wiping down the filter disk with alcohol, replace the lid. Each time I do an injection the needle is sterilized and handled with fresh gloves. For some reason this has consistently resulted in a sour patch contamination.
It sounds like your LC is contaminated. This is exactly the reason I recommend against LC. You never know what you have until six weeks later, and then if it's bad, you're back to square one. Use spores or learn agar skills, but LC is not going to give a good percentage of success compared to other methods. RR
Day 4 bump of the experiment. Knocked up the jars on the 18th, and here's the update:
In the martha, the ecuador and burma LC injected jars are showing white mycellium. The direct spore injected jar is showing nothing yet. On the ambient temp shelf there's growth in the ecuador LC jar and the others are showing nothing. The control jars in both environments are still clean thus far. Predictably this growth has shown signs of stopping after about 25% colonization in previous attempts, so I'm not too optimistic yet. I'll leave my jars unshaken until I see a 50% colonization. Typically I start to see the moisture buildup indicating a wet spot/sour patch infection after about a week. More updates will be forthcoming.
In the meantime, my mind's reaching for alternatives to fight the sour patch. If the acidity lowers the ph of the grain to make them uncolonizable, would adding a basic solution(such as baking soda, an antacid, or even ground lime) to the soaked grains before PCing help my myc get a handup on colonization before the sour patch can get a foothold?
I am seeing signs of white mycellium in all of the jars that were purposely inoculated, save those injected directly with spores. The Burma is definitely growing at least 30% faster than the Ecuador, both in the Martha and on the shelves.
The jars within the Martha are visibly cultivated roughly 10%, give or take. As of yet there is no sign of any contamination in the Control Jar, or in any of the jars.
On the ambient temperature shelf the LC Burma and LC Ecuador jars have cultivated roughly 5%. In the Burma jar I have noticed a small amount of light beige-colored fluid, slightly opaque in color roughly the size of a dried split pea attached to the inner glass of the jar, forming in a particularly dense patch of mycellium roughly the size of a quarter. I have noticed no off odor, and thus I suspect it to be an excretion of a relative metabolic compound produced by the mycellium. I will closely observe for signs of reduced growth. On the Ecuador LC jar I detect no signs of contamination, but something worthy of note is that the mycellium is a denser, and thusly brighter white. The Control Jar is visibly contaminant free.
I'm learning a lot from this experiment in more ways than I expected. Notably the differences in the rate of cultivation affected by strain and temperature. I am also theorizing that perhaps if the LC is not contaminated, perhaps the spawn bags may have come into contact with wet spot spores prior to my filling with prepared seed. It stands to reason that as the bags were not cleaned before filling, there may have been remnant endospores present that came into contact with them. Either washing the bags with a peroxide and phosphoric acid solution will fix the problem, or doing the same with jars will prevent further contaminations. I will not consider this a solution or conclusion until more time has passed, however.
The burma jars all have appeared to be roughly 50% cultured, so I gave them all a good shaking this morning. Also the ecuador spores directed straight from syringe have begin germinating. Still no sign of contamination.
At day 13 I feel ready enough to come to a conclusion on my experiment.
The issue with contamination that I sought to isolate was my spawn bags. They were the only thing that was changed in the experiment, and all samples are completely uncontaminated. If I use bags in the future I will make an effort to thoroughly sanitise them via moistening and soaking before filling with seed prior to PC sanitation. Or to curtail the extra step, I will simply stick with my jars.
Nearly all of my jars(including those inoculated with spores) are culturing happily away and are between 80 and 100% cultured. Tomorrow I'll begin adventures in spawning to bulk.
The lessons I learned from this are many, as are my new findings.
Ecuador spores injected from syringe will germinate faster at ambient room temperature than at 80-85 degrees, however once germinated the rate of growth equalises, as the mycellium spreads faster in a temperature of between 80-85 degrees. When injecting Ecuador from spores, save your energy and grow on a shelf.
When cultured from LC, Burma grows as much as three times faster than Ecuador in a heated environment kept between 80-85 degrees at all times. To counterpoint this, the Ecuador LC cultured it's jar on the ambient shelf almost as fast as the Burma did in the warm environment. Thus, Ecuador favours more ambient temperatures for culturing than Burma, which is suited to a more tropical ambient temperature.
Agreed, it's probably the LC. I had the same problem. I thought it was my procedure, but a few weeks later I noticed my LC wasn't quite right. I'll post a picture of my LC tomorrow, cause I haven't gotten to throwing it out yet. Should be pretty gnarly by now and that's at 68 degrees farenheit.
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