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OfflineLTSwoomz
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How am I still getting contams?
    #18932610 - 10/04/13 06:39 PM (10 years, 3 months ago)

I've been having a horrible time getting WBS to colonize. I got a new syringe and whipped up a new batch of WBS. Knocked them up on the 30th. Today I found it contaming. Or at least, that's what I think it is.



Both jars are like this.

WBS prep is Frank's. SHIP and SFD lids. PC'd at 15psi for 90m. Prepped my SAB like one of Frank's guides, paper towel damp with soap in it, wiped the whole thing down on the inside, let the puddle of soapy water sit. Put jars and flame sterlized need inside. Knock them up, move them to their resting space. I don't see where I went wrong this time.


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Edited by LTSwoomz (10/04/13 06:43 PM)


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InvisibleSmeagol
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932655 - 10/04/13 06:49 PM (10 years, 3 months ago)

Looks like my myc. From reading we are the biggest contamination risk. How do you prepare your grain? Do you use a still air box? how much solutIon dId you use? You need to give specific details to get specific answers.

Nevermind thought the details were you Sig. Derp de Der


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Edited by Smeagol (10/04/13 06:51 PM)


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: Smeagol]
    #18932688 - 10/04/13 06:54 PM (10 years, 3 months ago)

Quote:

Smeagol said:
Looks like my myc. From reading we are the biggest contamination risk. How do you prepare your grain? Do you use a still air box? how much solutIon dId you use? You need to give specific details to get specific answers.

Nevermind thought the details were you Sig. Derp de Der




1cc solution per jar. It's a SAB with gloves built in.

Doesn't the bottom of the jar look bad though? Or does it look normal?


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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932703 - 10/04/13 06:58 PM (10 years, 3 months ago)

That is a very large question.

The answer is either spores, inoculation technique, or grain prep.

There are several avenues that can be ventured under any one of those. Unfortunately, there is not enough info to give a well reasoned answer.

I don't trust spores from any source. That's why I use agar first.


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: SpitballJedi]
    #18932796 - 10/04/13 07:24 PM (10 years, 3 months ago)

How does one do agar without first having something to grow on it?

Does this look like contam to you?


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InvisibleEnigma1
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Re: How am I still getting contams? [Re: Smeagol]
    #18932798 - 10/04/13 07:25 PM (10 years, 3 months ago)

Are you doing the toilet paper test? Are you rinsing the WBS down good enough? I had to wash mine 3 times before it was right. Are you telling the bird seed how much you love  it everyday or are you telling it I know you are going catch a contam?


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InvisibleEnigma1
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Re: How am I still getting contams? [Re: Enigma1]
    #18932819 - 10/04/13 07:31 PM (10 years, 3 months ago)

If it was me I would isolate that batch and do some controversial experiments and see can you save it if it's a contam. I'm too new to tell you a answer with confidence.


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Offlinenn-IlliniSpiralDMT
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Re: How am I still getting contams? [Re: Enigma1]
    #18932820 - 10/04/13 07:31 PM (10 years, 3 months ago)

Usually contams from grain are from poor grain procedure in my experience. I like to make sure my grains are actually dry on the outside. Use a screen for a window as a strainer and leave it outside for 30 minutes. You'll have perfect grains:thumbup:

Grains should not be wet at all, like zero on the outside. You should feel that they are dry but when you squish them they should smash like a baked potato(only comparison I can think of lol).


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Re: How am I still getting contams? [Re: Enigma1]
    #18932826 - 10/04/13 07:32 PM (10 years, 3 months ago)

Do you let your pressure cooker expel all of the air before putting the weight bell on? If you don't well you aren't steam sterilising properly.

I would agree with Spitball, the idea of spores to growing medium makes me cautious. Are you interested in getting into working with agar? Now maybe the time to start.


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: nn-IlliniSpiralDMT]
    #18932829 - 10/04/13 07:32 PM (10 years, 3 months ago)

I've been doing a ton of WBS lately and I've made 100% sure these guys are dry on the outside, they pass the paper towel/toilet paper test everytime.


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932837 - 10/04/13 07:34 PM (10 years, 3 months ago)

I'm doing WBS for bulk growing. I do want to get into Agar, but assumed I couldn't do it with just syringe spores.

My pressure cooker is electric, it starts the countdown automatically when it reaches 15 psi.


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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932855 - 10/04/13 07:37 PM (10 years, 3 months ago)

You need to ensure the air is forced out of the PC for about 10 minutes for an effective sterilisation. I dont know if any automated system can do that.


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InvisibleEnigma1
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932857 - 10/04/13 07:38 PM (10 years, 3 months ago)

Quote:

LTSwoomz said:
I'm doing WBS for bulk growing. I do want to get into Agar, but assumed I couldn't do it with just syringe spores.

My pressure cooker is electric, it starts the countdown automatically when it reaches 15 psi.



When my pressure cooker is done I put a alcohol soaked towel over the vent.


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: barong]
    #18932865 - 10/04/13 07:40 PM (10 years, 3 months ago)

Quote:

barong said:
You need to ensure the air is forced out of the PC for about 10 minutes for an effective sterilisation. I dont know if any automated system can do that.




I've no idea how to go about doing that. The instructions for the PC are to set the vent to seal, turn the PC to how long you want it to run for at High setting (15 psi, I looked up this model) and it pressure cooks it.


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Invisiblebarong
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932875 - 10/04/13 07:44 PM (10 years, 3 months ago)

Your PC doesn't have a pressure regulator? Tell me what model you have.

In the meantime, here's a post I found by one of this forums well regarded growers, Roger Rabbit.

http://www.shroomery.org/forums/showflat.php/Number/9357332#9357332


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: barong]
    #18932882 - 10/04/13 07:46 PM (10 years, 3 months ago)

Ok give me about 45m and ill get that model number for you.


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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932900 - 10/04/13 07:51 PM (10 years, 3 months ago)

No worries. Not saying that this will be the answer to your issues, but its always good to ensure best practice in every part of your procedure.

Another thing, are you hydrating your grain correctly? With either WBS or wheat I have found that dry:hydrated tends to be around 1:1.6

Or in other words, if you start off with a kilo of grain, after you have hydrated, you should have 1.6 kilos.

Can you talk us through your grain prep in detail?


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Re: How am I still getting contams? [Re: LTSwoomz]
    #18932980 - 10/04/13 08:10 PM (10 years, 3 months ago)

Quote:

LTSwoomz said:
How does one do agar without first having something to grow on it?

Does this look like contam to you?





Quote:

LTSwoomz said:
I do want to get into Agar, but assumed I couldn't do it with just syringe spores.




Spore syringes are a good reason to get in to agar.

Use your spore syringe to put a small drop of spores on it. This will germinate and colonize. Just like in your grain jars.

If there is no contamination, then it's your grain jar prep.

If your agar gets a contamination, then you will have isolated the problem down to either inoculation technique or spores.

Either way, on agar, you can separate the good mycelium from the contam and put it on another agar dish. You can keep isolating and transferring until you no longer see a contamination.

If you keep getting contamination on your agar dishes, then the problem is your technique.

If your contam is gone after 1 or 2 transfers, then the problem is your spores.

This process is cheaper and faster than continuing to prep grains and waiting for colonization. One spore syringe can be used on many many agar dishes because it on takes a drop of spores. You could get your problem identified with less than 1-2cc's of spore solution. If you are astute, then you can do it with less than 1cc.


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: barong]
    #18933000 - 10/04/13 08:16 PM (10 years, 3 months ago)

WBS prep is as follows.

Grain is rinsed and strained of all floaters and sunflower seeds. I do this repeatedly until water runs clear. Gypsum is added before the rinse if I remember to add gyspum. After the rinse, I leave it to soak for ~18 hrs. Next day I bring it to a boil and immediately take it off of the stove and strain it with a net colander. It's shaken every 10m or so for 90m and always passes the paper towel test.


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: SpitballJedi]
    #18933008 - 10/04/13 08:17 PM (10 years, 3 months ago)

Quote:

SpitballJedi said:
Quote:

LTSwoomz said:
How does one do agar without first having something to grow on it?

Does this look like contam to you?





Quote:

LTSwoomz said:
I do want to get into Agar, but assumed I couldn't do it with just syringe spores.




Spore syringes are a good reason to get in to agar.

Use your spore syringe to put a small drop of spores on it. This will germinate and colonize. Just like in your grain jars.

If there is no contamination, then it's your grain jar prep.

If your agar gets a contamination, then you will have isolated the problem down to either inoculation technique or spores.

Either way, on agar, you can separate the good mycelium from the contam and put it on another agar dish. You can keep isolating and transferring until you no longer see a contamination.

If you keep getting contamination on your agar dishes, then the problem is your technique.

If your contam is gone after 1 or 2 transfers, then the problem is your spores.

This process is cheaper and faster than continuing to prep grains and waiting for colonization. One spore syringe can be used on many many agar dishes because it on takes a drop of spores. You could get your problem identified with less than 1-2cc's of spore solution. If you are astute, then you can do it with less than 1cc.




Can agar work be done successfully in a SAB then?


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InvisibleSpitballJedi
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18933014 - 10/04/13 08:18 PM (10 years, 3 months ago)

Your jars don't look too wet to me. If the problem is your grain prep, I would say it's either the sterilization process or GE filter.


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InvisibleSpitballJedi
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18933034 - 10/04/13 08:22 PM (10 years, 3 months ago)

Quote:

LTSwoomz said:
Can agar work be done successfully in a SAB then?




Absolutely. Myself and many people use a SAB for agar work.

I prefer not having the gloves attached though. That way I can flame sterilize outside the SAB and easily get my hand back in.

I know what you are going to ask, and no, this does not increase the risk of contamination. Many people can attest to it.


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Invisiblebarong
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Re: How am I still getting contams? [Re: SpitballJedi]
    #18933177 - 10/04/13 08:58 PM (10 years, 3 months ago)

Another one here who uses an SAB without gloves for agar work.

Your grain prep sounds solid by the way.


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: SpitballJedi]
    #18933287 - 10/04/13 09:24 PM (10 years, 3 months ago)

Quote:

SpitballJedi said:
Quote:

LTSwoomz said:
Can agar work be done successfully in a SAB then?




Absolutely. Myself and many people use a SAB for agar work.

I prefer not having the gloves attached though. That way I can flame sterilize outside the SAB and easily get my hand back in.

I know what you are going to ask, and no, this does not increase the risk of contamination. Many people can attest to it.




The GE filters are SFD's siliconed on. Not sure of any better ways to do GE holes.

Would you mind detailing your SAB procedure? How do you make sure the air inside stays still while you work, and what precautions do you take before and during to prevent contams from floating into your work?

Quote:

barong said:
Another one here who uses an SAB without gloves for agar work.

Your grain prep sounds solid by the way.




Good to hear. I'll definitely be reading into this.


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Edited by LTSwoomz (10/04/13 09:32 PM)


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InvisibleFrankHorrigan
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18933695 - 10/04/13 10:51 PM (10 years, 3 months ago)

Quote:

LTSwoomz said:
Can agar work be done successfully in a SAB then?




Yes :lol:


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: FrankHorrigan]
    #18940044 - 10/06/13 12:28 PM (10 years, 3 months ago)

So is this for sure contamination? I'm not just mistaking early myc for contam am I?


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Re: How am I still getting contams? [Re: LTSwoomz]
    #18941798 - 10/06/13 06:55 PM (10 years, 3 months ago)

Quote:

LTSwoomz said:
So is this for sure contamination? I'm not just mistaking early myc for contam am I?




That is for sure contamination.

Curious- was it from a vendors syringe or did you make the syringe yourself from a print?


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: Gretchenmeister]
    #18941893 - 10/06/13 07:09 PM (10 years, 3 months ago)

Quote:

Gretchenmeister said:
Quote:

LTSwoomz said:
So is this for sure contamination? I'm not just mistaking early myc for contam am I?




That is for sure contamination.

Curious- was it from a vendors syringe or did you make the syringe yourself from a print?




Vendor.


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OfflineRogerRabbitM
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18942077 - 10/06/13 07:50 PM (10 years, 3 months ago)

Quote:

LTSwoomz said:
So is this for sure contamination? I'm not just mistaking early myc for contam am I?




When it's anything but white early it's contamination.

You need your pressure cooker to vent steam for a few minutes before it seals up to build pressure.  If that's happening, they you're OK.  If it's sealed from the start, forget it.  It might cook food that way, but won't sterilize.

That said, a blue or green color is mold, and molds are killed at a temp below boiling. I suspect it's your inoculant at fault, since all the mold is at the bottom of the jar where that stuff ran.  If it was the filter, the mold would be on top, and if it was due to bad sterilization, the mold would be throughout the jar.
RR


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OfflineLTSwoomz
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Re: How am I still getting contams? [Re: RogerRabbit]
    #18942266 - 10/06/13 08:38 PM (10 years, 3 months ago)

Hm. Well these are brand new syringes. Suppose to solve a problem like this, I would need to start doing agar. I don't really know of any way other than that to make sure my inoculant is good.

And I will start letting my PC steam for 10m before closing the valve from now on.


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Edited by LTSwoomz (10/06/13 08:39 PM)


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Invisiblebarong
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Re: How am I still getting contams? [Re: LTSwoomz]
    #18942454 - 10/06/13 09:36 PM (10 years, 3 months ago)

Quote:

LTSwoomz said:
And I will start letting my PC steam for 10m before closing the valve from now on.




:thumbup:

And try and keep everything in your room as clean as possible when you're inoculating. I spray down all the work areas several times with spirits, also the air in the room. Try to do everything first thing in the morning when your house has had little air turbulence to blow the dust around, and go straight from the shower to the work area either in clean clothes, or often simply in the nude. Wash down your hands and arms with alcohol or a 10% bleach solution, don't breathe , cough or sneeze over your work...... be mindful of all of these things, each of which may not be a big deal but it's a good practice to get into.


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