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Offlinecamwill
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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.


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InvisibleMad Season
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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.


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Edited by Mad Season (07/18/15 06:44 AM)


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Offlinecamwill
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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


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InvisibleMad Season
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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


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Edited by Mad Season (07/18/15 07:15 AM)


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Offlinekn33b
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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.


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Offlinespacechildo
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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..


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Offlinecamwill
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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?


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Offlinelovesquare
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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.


--------------------
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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,
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Come back to the love that you had here with me...


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InvisibleJuiceh
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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.


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RogerRabbit said:You need a bigger pressure cooker for this hobby. RR


Edited by Juiceh (07/18/15 09:09 PM)


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Offlinecamwill
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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!!!


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InvisibleJuiceh
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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.


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OfflineMachiavelliavore
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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)


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Invisibleblindingleaf
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Re: Need help with grain spawn!!! [Re: Machiavelliavore]
    #21986404 - 07/23/15 05:50 PM (8 years, 6 months ago)

Quote:

5 micron will let alot of nasties in.




:confused:

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.


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Offlinecamwill
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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?


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InvisibleMad Season
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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


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How to shroomery like a pro! (Seriously, everyone read this!)
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AMU Q&A
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Offlinecamwill
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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?


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InvisibleMad Season
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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.


--------------------
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How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
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Offlinecamwill
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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.


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InvisibleJuiceh
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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.




:confused:

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.


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How I make 15 lbs of casing in a 941. How I line 12 shoeboxes with 1 39gal bag. How I Deep Fry Cubes!
RogerRabbit said:You need a bigger pressure cooker for this hobby. RR


Edited by Juiceh (07/23/15 08:18 PM)


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InvisibleMad Season
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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.


--------------------
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How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


Edited by Mad Season (07/23/15 08:14 PM)


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