Home | Community | Message Board


High Mountain Compost
Please support our sponsors.

Mushrooms, Mycology and Psychedelics >> Contamination Forum

Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Amazon Shop for: ½ Pint Jars, Autoclave, Impulse Sealer, Wild Bird Seed

Jump to first unread post. Pages: 1
InvisibleChibboleth
member
Registered: 10/14/02
Posts: 232
Chronic bacteria problems; what can be done?
    #2465484 - 03/23/04 11:30 PM (12 years, 8 months ago)

My foaf has attempted four grows in the past, with varying degrees of success. He is now on his fifth, which is his first using whole grains. Here's the thing: on every single grow he has at least one or two jars develop bacterial infection during incubation. This time it looks like no exception, and he was damn careful about PCing and inoculation.

He never gets mold at this stage. Any jar that makes it to birthing is almost always good for two or three flushes. What might be causing this chronic bacteria during incubation?

For example, this time:

Millet soaked for fourteen hours, PC'ed for 75-80 minutes in jars with polyfill lid filters. Prior to inoculation (in well-lit kitchen) all nearby surfaces wiped down with clorox disinfectant wipes. Aerosol lysol sprayed liberally around area. FOAF held his breath during inoculation itself, and flamed the syringe needle with a butane lighter between jars. Did he miss anything there?


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineSuntzu
Geek
 User Gallery

Registered: 10/15/99
Posts: 1,394
Last seen: 30 days, 18 hours
Re: Chronic bacteria problems; what can be done? [Re: Chibboleth]
    #2465755 - 03/24/04 12:35 AM (12 years, 8 months ago)

Bacterial contamination of whole grain jars is more often a result of incomplete sterilization and/or improper cooling.  If improper cooling was to blame, you'd expect a percentage of the contamination to be mold.  This isn't a hard and fast rule, but what is?  :wink:

There is no specific reason I can think of why pre-soaking would be a problem, and it sounds like the sterilization time was long enough [perhaps too long?].
I would check to make sure the moisture content is correct;  Overly wet jars will tend to clump at the bottom, and produce 'burst grains'.  Burst grains are more conducive to bacterial proliferation.  If s/he is confident about the moisture level, you may want to try an alternative grain prep.  The many that exist here are all good; however success of a tek varies from person to person. 
I think that regardless of the method, it's a good idea to rinse millet/birdseed pretty extensively before loading into jars.  I believe it cuts down on the endospore load.  For my gourmets, I never pre-steep/soak, but load the jars with rinsed grain and add an appropriate amount of water.  In this tek:

http://www.shroomery.org/forums/showflat...rev=#Post644625

the water level used is higher than most should use, but the mix is more milo than millet.  [cheap ass birdseed].

You may want to consider telling your friend to reduce the 15 PSI time to an hour, it should be enough. . .unless no burst grains have been seen?

The other potential problem might be in the inoculation;  simple gloveboxes can really make a cleaner environment, but again--I'd expect to see at least the occasional mold contam if that were to blame. . .here's one that is cheap and works well;



Best of luck!

**also, another thing to consider is the inoculum itself.  Spore solutions/prints can be contaminated. . .negative controls can help identify this problem.


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineShroomInhaler
No, I don'tsmoke them! :P

Registered: 03/04/04
Posts: 102
Last seen: 9 years, 8 months
Re: Chronic bacteria problems; what can be done? [Re: Chibboleth]
    #2465758 - 03/24/04 12:37 AM (12 years, 8 months ago)

Suntzu beat me to it and said all that needed to be said. :smile:


Edited by ShroomInhaler (03/24/04 12:49 AM)


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleChibboleth
member
Registered: 10/14/02
Posts: 232
Re: Chronic bacteria problems; what can be done? [Re: Suntzu]
    #2466711 - 03/24/04 09:31 AM (12 years, 8 months ago)

Quote:

Suntzu said:I would check to make sure the moisture content is correct; Overly wet jars will tend to clump at the bottom, and produce 'burst grains'. Burst grains are more conducive to bacterial proliferation.



There are some burst grains, yes. Just to be clear - if the contents of the jar are actually sterile, why would burst grains be a problem? Or are they some kind of haven for the odd bacterium that manages to get through the polyfill? Still trying to narrow down the source to either "substrate" or "environment". Jars were cooled in an oven, by the way.

As for contaminated inoculum... doubt it. This was a Ralphster's syringe, and he took great care in the process of attaching the needle to the syringe. Plus if it was that I'd expect all the jars to have problems instead of just one (and it's even possible that the one is alright - it might just be soggy kernels. The appearance isn't conclusive yet.).


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineSeussA
Error: divide byzero

Folding@home Statistics
Registered: 04/27/01
Posts: 23,480
Loc: Caribbean
Last seen: 25 days, 22 hours
Re: Chronic bacteria problems; what can be done? [Re: Chibboleth]
    #2466749 - 03/24/04 09:54 AM (12 years, 8 months ago)

I had a similiar problem when I first switched to grain. I would increase my soak times, increase my PC times, increase my pressures (I was using an autoclave), and nothing seemed to work. I tried different syringes from different sources, different strains, etc. Every single bag of grain would still turn into a bag of slimey goo a week or two after innoculation

I eventually located the source of my problems. My incubator (real incubator, not a do-it-yourself type) was contaminated. I am not sure how those little bastards managed to get into my grain, but they did. I was using bags with filter patches sealed with an impulse sealer. I ended up soaking all of the shelves in my inncubator in hot bleach water for several hours, rinse, and then soak again. I wiped out the inncubator with bleach, then alcohol, then bleach again. Finally I put it all back together and "set off a can" of lysol inside the incubator and let the insides soak in lysol fumes for a few days. After that I had no more problems with bacteria.


--------------------
Just another spore in the wind.


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineSuntzu
Geek
 User Gallery

Registered: 10/15/99
Posts: 1,394
Last seen: 30 days, 18 hours
Re: Chronic bacteria problems; what can be done? [Re: Chibboleth]
    #2466751 - 03/24/04 09:55 AM (12 years, 8 months ago)

You're right, if the contents are sterile, burst grains should make no difference.
But it seems to, IME. 
The inside of the seed is the starchy part;  burst seed provides a high surface area/starch content which seems more bacteria-friendly.  The almost-waxy coat of millet is very smooth;  less surface area, less free starch. . .mycelium make better ground on it.  The times I've tried steeping or pre-soaking pure millet, it has burst.  This doesn't happen for other people, though.
Something that Chibboleth alluded to [and perhaps played a role for Seuss?]. . .if the entire inside of the jar is coated with starchy residue [from burst grain], this may facilitate the 'wicking' action of bacterial contams.  Bacillus are motile bacteria, and if there is a moisture/nutrient path, they could potentially maneuver even around a polyfill plug and 'swim' down into the insides.  Non-burst grains don't produce as much of this. . .it can look like 'slime' on the sides of a jar after shaking.
I'm sure the inoculum is fine, too;  :smile:


Post Extras: Print Post  Remind Me! Notify Moderator
Jump to top. Pages: 1

Amazon Shop for: ½ Pint Jars, Autoclave, Impulse Sealer, Wild Bird Seed

Mushrooms, Mycology and Psychedelics >> Contamination Forum

Similar ThreadsPosterViewsRepliesLast post
* Bacteria problem?? (pics) Olgualion 1,283 7 07/22/02 08:26 AM
by Olgualion
* bacteria in g2g jars 0003 935 4 07/18/06 11:07 PM
by RogerRabbit
* More Lost Jars Anonymous 654 1 06/16/02 07:50 AM
by MAIA
* 90% colonization, some bacteria artemus 1,418 2 11/19/02 05:22 PM
by Anonymous
* Burst grains and bacteria MushBus 991 3 11/02/04 02:36 PM
by MAIA
* problems with bacterial contams in g2g 0003 1,535 11 08/21/06 03:58 PM
by monstermitch
* Problems with LCs... beatnicknick 1,158 6 07/13/06 02:59 AM
by beatnicknick
* Jars harboring bacteria ke1n 1,414 4 01/14/07 12:01 AM
by RogerRabbit

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: RogerRabbit, cronicr, Pastywhyte
1,148 topic views. 2 members, 4 guests and 3 web crawlers are browsing this forum.
[ Toggle Favorite | Print Topic | Stats ]
Search this thread:
Azarius
Please support our sponsors.

Copyright 1997-2016 Mind Media. Some rights reserved.

Generated in 0.051 seconds spending 0.003 seconds on 14 queries.