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Shop: Mushroom-Hut Liquid Cultures   North Spore Injection Grain Bag

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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
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Funky smell after only four days in the incubator
    #2141016 - 11/27/03 08:25 PM (20 years, 4 months ago)

A friend of mine innoculated 8 brf jars on the 23rd, four days ago. He put them in his makeshift incubator, a 10 gal aquarium filled with salt water and a 50 watt heater. Much to his dismay, by the third day, there was a rank, salty smell to the entire aquarium. He pulled out each of the eight jars and smelled them, finding that each jar had the same smell. He figured that he'd put too much salt in the aquarium, since after four days, it was very unlikely that each jar had been contaminated with the exact same infection. He removed and carefully inspected each of the jars, and since he saw nothing out of the ordinary, figured it was the salt content of the aquarium. He picked up the aquarium to take it to the bathroom for a thorough disinfecting, when he realized that once the jars were gone, the smell had disappeared from the aquarium. Clearly it was the jars making the smell. He was upset. How could bacteria have infected each of the eight jars in a little under four days?!? The incubator, while not ideal, shouldn't have caused this much trouble. Each of the eight half pint jars was pc'd for more than 45 minutes at 15 psi. What gives?


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InvisibleShroomismM
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Re: Funky smell after only four days in the incubator [Re: turtlelamp]
    #2141519 - 11/28/03 12:57 AM (20 years, 4 months ago)

Why salt? Put a little hydrogen peroxide in the water next time


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OfflineMAIA
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Re: Funky smell after only four days in the incubator [Re: Shroomism]
    #2141671 - 11/28/03 05:19 AM (20 years, 4 months ago)

The jars, were they in direct contact with the water ?
Anyway, you shouldn't use salt for this purpose. Like shroomism said, use h2o2. I used to add a little bit of bleach, which is also very good.

MAIA


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OfflineSuntzu
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Re: Funky smell after only four days in the incubator [Re: turtlelamp]
    #2142011 - 11/28/03 11:12 AM (20 years, 4 months ago)

IMO the jars were contaminated before incubating. When all jars fail like this the simplest explanation is either;

a) They were incompletely sterilized and/or improperly cooled
b) Innoculant/Innoculation step was to blame

How high on the jar did the water level of the incubator go? Was it high enough to potentially 'wick' bacteria around the neck of the jars?

To really get to the bottom of this problem, a couple of negative controls would be helpful. One that is uninnoculated and incubated with the others. . .One that is 'fake innoculated' [needle inserted, but no fluid injected] and incubated with the others.

It's possible that the offender originated in the incubator, but the vast majority of the time it's from elsewhere.

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Invisibleturtlelamp
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Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Funky smell after only four days in the incubator [Re: Suntzu]
    #2142100 - 11/28/03 12:11 PM (20 years, 4 months ago)

Bleach is a good idea. As far as h2o2, I don't think that would be as effective, since even small amounts of light cause it to rapidly break down. I can't imagine that his sterility measures are to blame... I mean, they probably are, but it's almost inconceivable. Perhaps the syringe is to blame after all. I'll tell him to use controls next time he prepares jars. As far as cooling, is there perhaps something he's missing? All he has been doing is pc'ing at 15 psi for 45-60 minutes and cooling overnight. Thank you all for your help and suggestions.


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