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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
Loc: The Midwest
Confused: Last three sets of jars have been failures
    #2161164 - 12/05/03 11:24 AM (20 years, 3 months ago)

I began about a month ago, and innoculated 6 half pint jars after only boiling them in a large pot. They all germinated and colonized. Of the next set of 6 I prepared, only one colonized. The next time I prepared 8 jars and pc'd them in a new cooker. Not one of those jars germinated. Last week I prepared 8 more and again, none of those germinated. I originally kept the jars in an aquarium w/heater where they were in direct contact with the water but now I have a setup similar to the one illustrated in pf-tek for simple minds. I've tried this so many different ways, using different amounts of water in the jars, keeping them at different temperatures, etc, and nothing is working. I'm not an idiot. And why did it work the first time when I was stumbling through it? I'm beginning to think that the spores I ordered from sporebank (I know, not a sponsor = bad news) are bad, but I checked closely at my last syringe and I could see the spores fairly clearly. Any thoughts? Have any of you had this many problems with getting your jars to take?


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Invisiblemicro
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2161183 - 12/05/03 11:28 AM (20 years, 3 months ago)

Could be the syringes. If you see black, though, usually they are good. You let the jars cool before inoculating?

If only a few spores went in (i.e. if most were clumped in one part of the syringe) it could take longer to see growth.

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Invisibleturtlelamp
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Loc: The Midwest
Re: Confused: Last three sets of jars have been failures [Re: micro]
    #2161197 - 12/05/03 11:33 AM (20 years, 3 months ago)

I let them cool overnight these last few times.  With my last set of 8, all of them eventually became contaminated with what I can only assume is some kind of bacteria.  I'm running out of ideas.  :confused:


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Invisiblemicro
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2161257 - 12/05/03 11:51 AM (20 years, 3 months ago)

Oh -- if they smelled bad then it was contamination, and that's the problem.... They smelled bad?

What kind of substrate and how long did you PC it at 15 psi? If sterilization isn't it you need to clean, clean, clean....

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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Confused: Last three sets of jars have been failures [Re: micro]
    #2161270 - 12/05/03 11:55 AM (20 years, 3 months ago)

The batch prior to the current one did smell funky. I pc'd them at 15 psi for 45 minutes. The thing is, every single jar had the same garbage-y smell. Every single one. What are the odds of that? And I was very careful, flame and alcohol sterilizing the needle after each innoculation. The current batch hasn't yet shown any signs of contamination, I only pc'd them for 30 minutes this time, but there's still no growth.

EDIT: They're just half pint brf jars


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Edited by turtlelamp (12/05/03 11:57 AM)

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Offlinehavatampa
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Registered: 10/07/03
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2161591 - 12/05/03 01:10 PM (20 years, 3 months ago)

Get different syringes (Ralphster) and try baking your brf and verm in the oven at 350 F for about 35 minutes before using it for you mix. Also, boil you water to use in the mix. Get a spray bottle and fill it with rubbing alcohol. Spray that everywhere (all hard surfaces). Go back over sterility faqs and make sure you incubate your jars in a completely sterile place. Also, mist jars with alcohol everytime you check them and try to open the incubation container as little as possible.


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OfflineBoobylip
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Re: Confused: Last three sets of jars have been failures [Re: havatampa]
    #2161613 - 12/05/03 01:16 PM (20 years, 3 months ago)

Are you saying that they get contams?
Or that they are just sitting there with no growth??


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Invisiblemicro
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Re: Confused: Last three sets of jars have been failures [Re: Boobylip]
    #2161664 - 12/05/03 01:32 PM (20 years, 3 months ago)

Quote:

The thing is, every single jar had the same garbage-y smell. Every single one.




That's bacteria.  The first thing you need to do is sterilize the jars for at least an hour at 15 psi -- you should be doing this, anyway.  Also, you can use rubbing alcohol and rub it on the holes right before inoculating, too.

Other than that, clean :wink:

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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Confused: Last three sets of jars have been failures [Re: Boobylip]
    #2161677 - 12/05/03 01:34 PM (20 years, 3 months ago)

It's a little of both. The first set worked. Every jar colonized. The second set didn't, nothing colonized. No contams, nothing. The third set didn't colonize either, and every jar eventually became contaminated with bacteria. Regardless, though, it didn't colonize. Even if contaminated, ONE of the jars should have germinated a LITTLE. I sincerely doubt the bacteria ate every single spore with in three days of innoculation. This last batch is a week old. Nothing has shown. They don't smell peculiar yet... just like cooked brown rice flour. I'm baffled and ready to blame it on the syringes.


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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Confused: Last three sets of jars have been failures [Re: micro]
    #2161690 - 12/05/03 01:37 PM (20 years, 3 months ago)

An hour at 15 psi? I've read in other posts that you shouldn't cook it longer than 30 minutes for fear of destroying the nutrients in the brf. Some sources only recommend 15 minutes.


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OfflineBlue Helix
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2163326 - 12/06/03 12:58 AM (20 years, 3 months ago)

Where are the jars in relation to the heater? If jars are too close to the heater, they will get too hot, even if the bulk of the water is the right temperature. I had this happen to me in the past.

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Invisibleturtlelamp
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Re: Confused: Last three sets of jars have been failures [Re: Blue Helix]
    #2163356 - 12/06/03 01:31 AM (20 years, 3 months ago)

I have a two-rubbermaid setup now. The heater is a goodly distance.


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OfflineBlue Helix
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2163378 - 12/06/03 01:50 AM (20 years, 3 months ago)

Even in a two-rubbermaid setup, you can have the jars that are right above the heater get really hot while those far are fine. I have a two-rubbermaid set up now but I was having trouble with certain jars on the bottom until I raised the jars off the floor of the rubbermaid. I guess the jars right above the heater were getting too hot. I recently posted about my current incubator. You might want to do a search and see it.

I am really just guessing here, but you should never underplay temperature, especially too hot temperatures. A sustained temperature of say 95F, just a little warm, is really, really bad news for mycelium in the early stages of growth because it boosts bacterial growth like crazy--and there is always bacteria trying to take hold--while at the same time high temps stunt mycelium growth. In a matter of days, your grain will smell bad and stall out.

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OfflineAnnoA
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2163436 - 12/06/03 02:52 AM (20 years, 3 months ago)

>An hour at 15 psi? I've read in other posts that you shouldn't cook it
>longer than 30 minutes for fear of destroying the nutrients in the brf.

You can safely PC them for an hour without any "nutrients being destroyed"


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InvisibleMagash
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Re: Confused: Last three sets of jars have been failures [Re: Blue Helix]
    #2163467 - 12/06/03 03:27 AM (20 years, 3 months ago)

Put a small water pump in and you?ll get rid of the problem. It mixes the water so it doesn?t just heat up from the bottom up.



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OfflineBlue Helix
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Re: Confused: Last three sets of jars have been failures [Re: Magash]
    #2163611 - 12/06/03 06:15 AM (20 years, 3 months ago)

Yeah I think a small pump (called powerhead) will work fine. One thing you have to remember if you go that route is that some powerheads are too powerful and will heat the water all by themself too much. Just make sure the pump you use is less than say 3 watts or so otherwise the water will get too hot just from the pump. I had a small powerhead that was 8.5W from an old fish tank that just heated the water too much. That is why I went with an old PC fan in the main chamber instead. Either way the idea is the same. You are trying to mix up the heat. It's real important that you do.

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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Confused: Last three sets of jars have been failures [Re: Blue Helix]
    #2163712 - 12/06/03 08:10 AM (20 years, 3 months ago)

I actually have a pump already. My basement can be cold, so I needed it to keep the edges of the tank from dropping in temp too much. I've been monitoring the temp closely and it hasn't raised past 82F. I'm waiting for some new spores now. If these fail, I'll have exhausted all the easy answers and will have to totally reevaluate my jar preparation procedures.


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OfflineBlue Helix
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2163872 - 12/06/03 10:14 AM (20 years, 3 months ago)

Oh, I see. I thought you were a little greener from you first post. Yeah, I'd consider the spore supplier and the jar prep. There are so many ways to fail. One thing I noticed is that you have to be sober to do jars well. I had a friend who tried to do some once like I told him but never could get them quite right when he would light up some green.

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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Confused: Last three sets of jars have been failures [Re: Blue Helix]
    #2163886 - 12/06/03 10:22 AM (20 years, 3 months ago)

Quote:

Blue Helix said:
... One thing I noticed is that you have to be sober to do jars well...




That's a magnificent suggestion. I will try that.


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Invisibleturtlelamp
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Re: Confused: Last three sets of jars have been failures [Re: turtlelamp]
    #2171986 - 12/12/03 12:50 AM (20 years, 3 months ago)

Incidently, with that last batch, all of the jars I innoculated eventually became contaminated (bacterial, precisely underneath the innoculation points). None of the control jars were affected. Mystery solved. Never buy from Sporebank.

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