|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Need help with grain spawn!!!
#21959433 - 07/18/15 06:38 AM (8 years, 6 months ago) |
|
|
Well after my last attempt at doing a bulk spawn run, and its failures I decided to start all over from scratch. And I'm still getting the same results! New culture, everything!
So here's the rundown
1. Started fresh culture from spore on agar (was very successful) 2. Inoculated 1 pint jars with agar wedges (colonized very quickly) 3. Prepared grain for bulk spawn Rinsed grain, simmered grain, re-rinsed and dried, packed into filter patch spawn bags (4 quarts per bag), PC'd in 32qt PC two bags at a time for 90 minutes @15psi. 4. Bags were allowed to cool completely, over night! 5. Created grain liquid culture from previously colonized jar and inoculated spawn bags (20cc per bag)
Now here is the problem. 4 days later the bags show no sign of growth (ok not that abnormal) but when I squeezed some of the air through the filter patch I'm getting that hard cider smell. They (6 spawn bags) were colonizing in a cabinet in the dark at room temp (~75f).
Am I not sterilizing the grain long enough? Is there something severely flawed going on here? hahaha I'm just really baffled because I never had these sort of issues when I used jars, I just decided to switch because of the ease of cleanup.
Suggestions or experience would be much appreciated.
|
Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
|
Re: Need help with grain spawn!!! [Re: camwill]
#21959441 - 07/18/15 06:44 AM (8 years, 6 months ago) |
|
|
I'm a little confused. Did you make a grain liquid culture by inoculating with a clean agar wedge? What do you mean agar > 1 pint jars? Also you should be doing a minimum of 1 agar transfer to make sure it is clean. Process should be spores> agar> agar again for cleanliness > either lc or grains. You can from there do what you want. Lc should get tested on a few dishes of agar after you make it to make sure it's clean, and the grains can get either spawned or g2gd.
Also bags should really get pcd for 2-3 hours. 90 minutes is nowhere near sufficient.
Edited by Mad Season (07/18/15 06:44 AM)
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: Mad Season]
#21959453 - 07/18/15 06:50 AM (8 years, 6 months ago) |
|
|
The cultures on dishes was transferred to second dishes, then to 1 pint jars of sterilized grain which they colonized. I prepared syringes with sterile distilled water, injected it into the colonized jars and pulled back the mycelium rich water and immediately inoculated the spawn bags.
I was always a little suspicious of 90 minutes not being long enough. I have had such a high failure rate recently and now I'm almost certain that was the issue.
I'll be preparing a fresh batch of grain in the next few days. Updates to come
|
Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
|
Re: Need help with grain spawn!!! [Re: camwill]
#21959471 - 07/18/15 07:02 AM (8 years, 6 months ago) |
|
|
Instead of making syringes out of jars like that, do the same but to colonized agar dishes. It'll be WAY cleaner, or look up liquid inoculant. Anything on agar is best because you can see bacteria and other molds, whereas grains almost always have bacteria you can't see. It'll also help you skip a step :P
That's why I prefer to work with agar or liquids, and just leave grains for spawning.
This video comes to mind
Edited by Mad Season (07/18/15 07:15 AM)
|
kn33b
Stay golden, pony boy.



Registered: 03/16/15
Posts: 850
Last seen: 7 years, 10 months
|
Re: Need help with grain spawn!!! [Re: camwill]
#21959483 - 07/18/15 07:16 AM (8 years, 6 months ago) |
|
|
I went through the same thing. Soak the grain for 24hrs and PC for longer in a reliable pressure cooker.
--------------------
All written material by this individual is for entertainment purposes only. All events described or pictures displayed therein are ficticious and do not necessarily reflect the author's opinions or intentions.
|
spacechildo
proletarians rise up



Registered: 01/24/13
Posts: 19,243
Loc: Babylon
Last seen: 6 years, 4 months
|
Re: Need help with grain spawn!!! [Re: kn33b]
#21959887 - 07/18/15 10:11 AM (8 years, 6 months ago) |
|
|
4qt bags for 90 mins is recipe for disaster. we do 1qt jars for 90mins.
and why didnt you just inoc the bag with the pint jar? its a bit confusing..
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: spacechildo]
#21960517 - 07/18/15 12:59 PM (8 years, 6 months ago) |
|
|
don't you seal your bags while they're hot so they're "vacuum sealed"? how would i get the colonized grain into a sealed bag?
|
lovesquare
Love²

Registered: 06/04/15
Posts: 556
Last seen: 8 years, 4 months
|
Re: Need help with grain spawn!!! [Re: camwill]
#21961001 - 07/18/15 03:18 PM (8 years, 6 months ago) |
|
|
In detail, explain your grain prep please.
-------------------- If you go down round the bend in the river, You're gonna find a few changes been going down there. If you go down to the gas-powered flatland, Where most of the people just think that they're free, Remember the peace that you had on the mountain, Come back to the love that you had here with me...
|
Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
|
Re: Need help with grain spawn!!! [Re: camwill]
#21962403 - 07/18/15 08:59 PM (8 years, 6 months ago) |
|
|
Quote:
camwill said: packed into filter patch spawn bags (4 quarts per bag), PC'd in 32qt PC two bags at a time for 90 minutes @15psi.
2 4qt bags in a 32qt pc? Did you have more bags? Surely you could fit more in there, even if that's a typo and its a 23qt. You should be able to fit 4 in there. I can fit 4 6lb sawdust bags in my AA921.
Quote:
camwill said: Created grain liquid culture from previously colonized jar and inoculated spawn bags (20cc per bag)
Why didn't you just G2G that jar to the bags instead of adding another vector for contams?
Quote:
camwill said: when I squeezed some of the air through the filter patch I'm getting that hard cider smell.
It's probably not a good idea to squeeze air through the filter patch, they aren't very thick, and you can easily compromise a bag doing this.
Quote:
camwill said: don't you seal your bags while they're hot so they're "vacuum sealed"? how would i get the colonized grain into a sealed bag?
This raises alot more questions. Are you opening the PC in front of a flowhood? Do you have tyvek sleeves in the neck of the bags so that the air pulled in on cool down is filtered? The "vacuum seal" in reference to bags means how the neck of the bags with the tyvek sleeves semi seals itself in the PC so that the only route into the bag is through the tyvek.
I do a fuckton of bags. 10-20lbs of grain in XL bags, 6lb sawdust blocks, I even PC entire tubs worth of substrate in XL bags. They are not sealed before PCing since I use .2 micron filters. I use tyvek sleeves in the neck of the bag. The bags are inoculated via grain transfer in front of a flowhood, then impulse sealed so they have a nice plenum of clean air.
If you are using 5 micron bags, you can seal them before PCing. However, I don't think this filter is recommended for sterilizing grains. 5 micron will let alot of nasties in.
If you do not use tyvek sleeves in the neck of the bags you should move your PC in front of your flowhood for cooling, if you have one. This wouldn't hurt if you have sleeves in the necks either.
Edited by Juiceh (07/18/15 09:09 PM)
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: Juiceh]
#21985751 - 07/23/15 03:37 PM (8 years, 6 months ago) |
|
|
Quote:
Juiceh said:
Quote:
camwill said: packed into filter patch spawn bags (4 quarts per bag), PC'd in 32qt PC two bags at a time for 90 minutes @15psi.
2 4qt bags in a 32qt pc? Did you have more bags? Surely you could fit more in there, even if that's a typo and its a 23qt. You should be able to fit 4 in there. I can fit 4 6lb sawdust bags in my AA921.
Quote:
camwill said: Created grain liquid culture from previously colonized jar and inoculated spawn bags (20cc per bag)
Why didn't you just G2G that jar to the bags instead of adding another vector for contams?
Quote:
camwill said: when I squeezed some of the air through the filter patch I'm getting that hard cider smell.
It's probably not a good idea to squeeze air through the filter patch, they aren't very thick, and you can easily compromise a bag doing this.
Quote:
camwill said: don't you seal your bags while they're hot so they're "vacuum sealed"? how would i get the colonized grain into a sealed bag?
This raises alot more questions. Are you opening the PC in front of a flowhood? Do you have tyvek sleeves in the neck of the bags so that the air pulled in on cool down is filtered? The "vacuum seal" in reference to bags means how the neck of the bags with the tyvek sleeves semi seals itself in the PC so that the only route into the bag is through the tyvek.
I do a fuckton of bags. 10-20lbs of grain in XL bags, 6lb sawdust blocks, I even PC entire tubs worth of substrate in XL bags. They are not sealed before PCing since I use .2 micron filters. I use tyvek sleeves in the neck of the bag. The bags are inoculated via grain transfer in front of a flowhood, then impulse sealed so they have a nice plenum of clean air.
If you are using 5 micron bags, you can seal them before PCing. However, I don't think this filter is recommended for sterilizing grains. 5 micron will let alot of nasties in.
If you do not use tyvek sleeves in the neck of the bags you should move your PC in front of your flowhood for cooling, if you have one. This wouldn't hurt if you have sleeves in the necks either.
It is a 32 quart presto pressure cooker. I can fit 4 bags in there but it's a very tight squeeze. I'm still confused about this whole "tyvek" sleeve business. Maybe you have a link to a tek for this? I've never had these problems with spawn bags in the past. Also, How are you doing a grain to grain transfer to your spawn bags? I was under the impression you impulse seal the bags while they're hot out of the pressure cooker? wouldn't that mean breaking the seal on the bags to pour in the colonized grain?
I started another batch of grain 3 days ago, grain still putting off the fermented smell. This is becoming costly and annoying. They were pc'd for two and a half hours this time. I'm so frustrated!!!
|
Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
|
Re: Need help with grain spawn!!! [Re: camwill]
#21986283 - 07/23/15 05:25 PM (8 years, 6 months ago) |
|
|
You sure you have a 32qt PC and not a 23qt? I'm not finding 32qt prestos online, only 23qt. Fitting 4 bags makes sense if its a 23qt. I can get 4 of that size in my 21.5qt AA921.
Tyvek sleeve tek: http://www.mushroomvideos.com/ Click the sawdust\woodchip prep video and skip to 1:20 and you will see how the tyvek is used.
The bags will kinda semi seal around the tyvek after the cook. When you want to inoculate the bags(in front of your flowhood or in your SAB) you pull out the tyvek sleeve, dump the spawn into the bag opening and then impulse seal. This allows you to get a nice plenum of clean air in the top of the bag.
I PC my grain bags for 120 mins at 20psi.
|
Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
Posts: 3,038
Loc: The Sporetorn States
Last seen: 3 months, 20 days
|
Re: Need help with grain spawn!!! [Re: Juiceh]
#21986386 - 07/23/15 05:45 PM (8 years, 6 months ago) |
|
|
I think you PC'd your bags too long. We all know things evolve faster in the heat, which is why tropical diseases come from the jungle. You have probably created some kind of super-bacteria.
J/K. What everyone else said. PC considerably longer for bags of that size. Just 2/3 of a quart will run 60+, typically 90 minutes. Also, just use your whole pint of grain spawn for a grain2grain noc.
--------------------
I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
Edited by Machiavelliavore (07/23/15 06:05 PM)
|
blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
|
|
Quote:
5 micron will let alot of nasties in.

not sure on that…wouldn't be a top selling product if it did, yea?
u can make grain spawn in .5's. pirate swazey does it, and pre seals.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: camwill]
#21986615 - 07/23/15 06:37 PM (8 years, 6 months ago) |
|
|
Also I'm just curious. I read a lot of wbs teks where people just pre soak the grains for 24 hours, skip the simmer, and dry and pc. Is this an effective method or is simmering necessary?
|
Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
|
Re: Need help with grain spawn!!! [Re: camwill]
#21986644 - 07/23/15 06:43 PM (8 years, 6 months ago) |
|
|
Soak and simmering will get them to hold the most water content they can without bursting
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: Mad Season]
#21986868 - 07/23/15 07:32 PM (8 years, 6 months ago) |
|
|
Quote:
Mad Season said: Soak and simmering will get them to hold the most water content they can without bursting
So this whole business of simmering to release "bacterial endospores" is a waste of my time then?
|
Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
|
Re: Need help with grain spawn!!! [Re: camwill]
#21986893 - 07/23/15 07:38 PM (8 years, 6 months ago) |
|
|
Um I always thought the myth was you soak to get endos to germinate. I've never heard that one before. Honestly you're not going to get rid of endos EVER until you pc. Everything else said is just some strange made up speculation.
However you do want the grains to hold the most water they can without both bursting or over cooking. The latter of which will cause clumpy grains.
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: Mad Season]
#21987034 - 07/23/15 08:06 PM (8 years, 6 months ago) |
|
|
Quote:
Mad Season said: Um I always thought the myth was you soak to get endos to germinate. I've never heard that one before. Honestly you're not going to get rid of endos EVER until you pc. Everything else said is just some strange made up speculation.
However you do want the grains to hold the most water they can without both bursting or over cooking. The latter of which will cause clumpy grains.
Well tomorrow I'm going to get some fresh grains and try again with a g2g transfer instead of my previous method.
This is all so bizarre to me though. This only started happening when I switched to using Spawn bags as opposed to jars. And my sterile procedure was far from sterile back then.
|
Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
|
Re: Need help with grain spawn!!! [Re: blindingleaf]
#21987068 - 07/23/15 08:11 PM (8 years, 6 months ago) |
|
|
Quote:
blindingleaf said:
Quote:
5 micron will let alot of nasties in.

not sure on that…wouldn't be a top selling product if it did, yea?
u can make grain spawn in .5's. pirate swazey does it, and pre seals.
I said 5 micron not .5 and see below.
Quote:
hamloaf said: Trichoderma spores are .3 microns in size so if given the option, I'd go with .2 micron filter patch to ensure trichoderma spores can't fit between the micron size of the filter patch on the spawn bags I am using.
Quote:
camwill said:
Quote:
Mad Season said: Soak and simmering will get them to hold the most water content they can without bursting
So this whole business of simmering to release "bacterial endospores" is a waste of my time then?
Part of the reason for the simmer\boil at the end of the soak is also so that the exterior moisture on the grains can steam off, leaving the exterior dry and the grains free flowing.
Edited by Juiceh (07/23/15 08:18 PM)
|
Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
|
Re: Need help with grain spawn!!! [Re: camwill]
#21987075 - 07/23/15 08:13 PM (8 years, 6 months ago) |
|
|
Well how big are the bags? 4qts? 90 minute pc is the minimum for 1 qt jars. Bags minimum pc cook times is more like 150 minutes.
Edited by Mad Season (07/23/15 08:14 PM)
|
Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
|
Re: Need help with grain spawn!!! [Re: Mad Season]
#21987115 - 07/23/15 08:20 PM (8 years, 6 months ago) |
|
|
FWIW, I've PCd grain bags for 90 minutes and had no issue. HOWEVER, this was done at 20psi. For peace of mind I usually run my bags for 120 mins at 20psi anyway.
|
Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
|
Re: Need help with grain spawn!!! [Re: Juiceh]
#21987140 - 07/23/15 08:25 PM (8 years, 6 months ago) |
|
|
Yeah higher pressures reduces cook times. 5 extra psi doesn't sound like much, but that's a huge difference. Personally my pc at 15 psi seems to make pint and qt jars almost always completely sterile at 120 minutes. 90 minutes really doesn't cut it. Soon I'll be getting a better pc that'll be able to do higher pressures with different weights . I'd probably run bags for 2.5-3 hours at 15 psi.
|
blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
|
Re: Need help with grain spawn!!! [Re: Mad Season]
#21987180 - 07/23/15 08:32 PM (8 years, 6 months ago) |
|
|
my bad J, i read it like this
Quote:
grains. 5 micron
and saw the "." in front of the 5 ur right
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: Mad Season]
#21987273 - 07/23/15 08:56 PM (8 years, 6 months ago) |
|
|
I was using the medium Spawn bags from outgrow. I'm not site how many microns the filter patches are but I'm officially out of Medium bags and will be using up my stock of Large bags. Any pointers?
|
Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
|
Re: Need help with grain spawn!!! [Re: camwill]
#21989105 - 07/24/15 09:38 AM (8 years, 6 months ago) |
|
|
Quote:
camwill said: I was using the medium Spawn bags from outgrow. I'm not site how many microns the filter patches are but I'm officially out of Medium bags and will be using up my stock of Large bags. Any pointers?
Out-grow carries .2 micron or 5 micron. Did you order both kinds? If not, you can lookup previous orders on the out-grow page and see what you ordered.
I do large bags as I described previously. I highly recommend a large flowhood for working with big bags. I've G2G'd from one XL bag to other XL bags. But even with my 4'x2' hood it's a bit tricky. I been thinking of stacking another 4'x2' on top of it for some extra headroom. G2Ging a qt jar of colonized grains into a bag is much easier. I don't think I would try XL bags in a SAB, too big, I think manipulating such a large item in a SAB would stir up the air a bit too much, but I suppose it could be done with a qt jar if you are careful.
|
camwill
Alchemist



Registered: 09/29/14
Posts: 46
Last seen: 6 years, 2 months
|
Re: Need help with grain spawn!!! [Re: Juiceh]
#22019289 - 07/30/15 10:47 AM (8 years, 5 months ago) |
|
|
So, here are some updates! Normally I'm incredibly anal and I just throw away stuff that I suspect to be contaminated. Lately I've been so busy with work I totally forgot that they were in the cupboard. This morning I had some free time and figured I'd pitch them today and to my surprise I find this!



So far all bags are at "relatively" the same level of colonization. I haven't handled them to inspect for any suspicious mold contams but everything I can see looks like healthy mycelium.
only took like 8 days 
I also have a large bag with about 7'ish quarts of wbs in it that I pc'd for about 4 hours. I would have gone longer but I suspected I was running low on water and in fact melted the bottom bag to the sidewall. 
I spawned that with the tyvek sleeve method and went g2g from pint jars of colonized grain and it's running very nicely (pics to come!)
|
|