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Adden

Registered: 06/04/03
Posts: 39,201
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I dunno if anyone brought it up yet. I was inspecting the lids of some screw top pp5's that failed nearly immediately. I used a drill for my holes and the micropore got punctured by leftover jagged pieces. Carefully inspected my mini rounds and one almost got past me. Just a heads up I guess.
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Adden

Registered: 06/04/03
Posts: 39,201
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Re: Pasty Agar Tek [Re: Adden]
#22918077 - 02/18/16 01:14 AM (8 years, 7 days ago) |
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..by the way if you tri-fold a regular sized paper towel and fold it into 4's the thing practically locks in it. Quicker and standardized and easier to just cut them.
Edited by Adden (02/18/16 01:21 AM)
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YoShroomhead420
PSILOCYBIN MUSHROOM GURU


Registered: 11/26/15
Posts: 221
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Re: Pasty Agar Tek [Re: Adden]
#22918284 - 02/18/16 04:22 AM (8 years, 7 days ago) |
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-------------------- Shroomhead, at your service.
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spacechildo
proletarians rise up


Registered: 01/24/13
Posts: 19,243
Loc: Babylon
Last seen: 6 years, 5 months
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only 1 more to go, you can do it
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YoShroomhead420
PSILOCYBIN MUSHROOM GURU


Registered: 11/26/15
Posts: 221
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I wanted to be able to find this thread easily.
-------------------- Shroomhead, at your service.
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Adden

Registered: 06/04/03
Posts: 39,201
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Hey sorry for bumping three times but I've seen a lot of people asking about agar contams. It's hard to take pics through pp5's so maybe it will be of help.
Here's some bacterial morphology.

Lovate, undulate and entire are what I've commonly seen.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Pasty Agar Tek [Re: Adden] 1
#22920498 - 02/18/16 05:49 PM (8 years, 7 days ago) |
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Most of the bacteria you are going to see is probably from inoculate and going to be pretty hard to nail down.
Bacillus is going appear mostly through insufficient sterilization or from contamed grains being used for inoculate. Otherwise bacterial contamination via user error could be nearly anything IME.
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TravelAgency
The ongoing "wow"


Registered: 12/25/10
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Last seen: 1 year, 24 days
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As long as this topic is active- every MS syringe I've put to agar has had contams present (or my sterile technique wasn't good enough- that's a possibility, though all my PastyPlates (thank you again sir) that i used to clone via cultures never had contams show up on them. This may also account for some of my failures in PFTek earlier on.
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pablokabute
Hari ng Amag



Registered: 11/22/11
Posts: 5,178
Loc: rural ghetto
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Quote:
Pastywhyte said: Most of the bacteria you are going to see is probably from inoculate and going to be pretty hard to nail down.
Bacillus is going appear mostly through insufficient sterilization or from contamed grains being used for inoculate. Otherwise bacterial contamination via user error could be nearly anything IME.
If we have excess water in our agar after hardening... wouldn't that raise bacterial issues also? Just as how our grains get bacterial contams when overly wet? Any experiences with that?
--------------------
Fermented Mushrooms!! --- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1 'The second seal: “All CONTAMINATED things and events are unsatisfactory.”' "I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST." --Anonymous Guerilla, or is he..
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Quote:
pablokabute said:
Quote:
Pastywhyte said: Most of the bacteria you are going to see is probably from inoculate and going to be pretty hard to nail down.
Bacillus is going appear mostly through insufficient sterilization or from contamed grains being used for inoculate. Otherwise bacterial contamination via user error could be nearly anything IME.
If we have excess water in our agar after hardening... wouldn't that raise bacterial issues also? Just as how our grains get bacterial contams when overly wet? Any experiences with that?
Excess water is not the cause of the bacteria, it just helps it spread easier. If you have bacteria in a plate it's either insufficient sterilization, dirty inoculate, or poor procedure.
Same thing with grains really. Fact is we do not actually kill every last bacteria rod or endospore when we sterilize. If your jars are swimming in nutrition laden water then it's going to allow any bacteria present to bloom. But the bacteria didn't come from the water. It came from the grain.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Well, agar does get properly sterilized, unlike grain, that's a huge factor too.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Pasty Agar Tek [Re: Inocuole]
#22922946 - 02/19/16 12:11 PM (8 years, 6 days ago) |
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Quote:
Inocuole said: Well, agar does get properly sterilized, unlike grain, that's a huge factor too.
Have you had a look at bodhis microscopy pics of sterilized MEA broths? Use a lot of LC and you get used to seeing blooms explode weeks after it initially tests clean and fruits grown from it.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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That doesn't really... sound fun. I was under the impression liquids were easy to fully sterilize. Never had any blanks contam on me..
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Pasty Agar Tek [Re: Inocuole]
#22922994 - 02/19/16 12:17 PM (8 years, 6 days ago) |
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They are pretty easy to sterilize by comparison and I am sure often complete sterilization does occur. But until I develop microscope eyes I am going to treat things as if they are not guaranteed sterile. In the end I feel it comes down more to population levels than anything.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Also keep in mind that while we can see bacteria on the surface of agar, that is only after enough reproduction has occurred to make them visible. Another thing to consider is that bacteria cannot grow through agar. But that doesn't mean there is none trapped in the middle.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
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These seem like possibilities that almost no one would have any control over. I mean... it's a fine sauce for the back burner of your mind, for when stuff comes up and there's no other explanation but... The information doesn't really contain the means to defeat itself, so I feel like actively considering those things would just lead someone to a paranoid panic about all their plates. Good to know at any rate... Should I be considering upping my PC times for agar just a couple minutes? I already do like 30@15.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Pasty Agar Tek [Re: Inocuole]
#22923095 - 02/19/16 12:39 PM (8 years, 6 days ago) |
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If what you are doing is working then keep doing it. Its when things stop working that we should consider these other aspects.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Now that coir and agar are straightened out my only issues left are with grains. They're the only chaos factor anymore it feels like.
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Adden

Registered: 06/04/03
Posts: 39,201
Loc:
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Re: Pasty Agar Tek [Re: Inocuole]
#22923228 - 02/19/16 01:24 PM (8 years, 6 days ago) |
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Slowly bringing my PC up every 15 minutes (usually three times) and running 45 mins at 15psi seems to work for me but I get a lot of condensation. It's going to depend on what kind of PC, your agar mix, kind of stove etc (from what I understand, please correct if wrong).
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: Pasty Agar Tek [Re: Adden]
#22923234 - 02/19/16 01:26 PM (8 years, 6 days ago) |
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Quote:
Dys said: Slowly bringing my PC up every 15 minutes (usually three times)
What does this mean...?
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