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Offlinerunsfrombears
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Need help isolating cause of bacterial contamination.
    #23627254 - 09/08/16 09:15 PM (7 years, 4 months ago)

I'm getting insane amounts of bacterial contamination on my G2G xfers.
Since I moved into this new apartment my success rate has dropped drastically. It's the rate of bacteria that has skyrocketed.
My procedure hasn't changed in years. Wbs soak 15 mins in hot water, drain overnight, Pc for 60 mins, Clean glovebox with alcohol, wipe jars with alcohol, spray disinfectant, wait 20 minutes, do g2g.
I'm not having any extra problems with my inoculations.
Is it possible that my new apartment is the problem? I'm the first tenant since the renovation.
I've just never had such an extreme amount of failures. I've tried everything I can think of to isolate the problem and I'm hoping someone had an idea I haven't thought of.
Btw, they aren't going sour when I don't open them ... So I know it's not my sterilization process.


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Offline99.99
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Re: Need help isolating cause of bacterial contamination. [Re: runsfrombears]
    #23627367 - 09/08/16 09:53 PM (7 years, 4 months ago)

Me to i just cant get rid of it so i live with it and do this
Antibiotic agar , lots of colonized agar per bag mixed super super well upon inoculation, after that DONT SHAKE ore mix up containers
You should have enough colinized agar in the spawn it wont need a mix
My graines fully colonize in about five days and doesn't allow the bacteria a foothold


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OfflinePsilocyBen17
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Re: Need help isolating cause of bacterial contamination. [Re: 99.99]
    #23627610 - 09/08/16 11:34 PM (7 years, 4 months ago)

how are you inoculating your masters?


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Offline99.99
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Re: Need help isolating cause of bacterial contamination. [Re: PsilocyBen17]
    #23627620 - 09/08/16 11:42 PM (7 years, 4 months ago)

Quote:

PsilocyBen17 said:
how are you inoculating your masters?




Me ? In a heppa hood


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OfflinePsilocyBen17
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Re: Need help isolating cause of bacterial contamination. [Re: 99.99]
    #23627649 - 09/09/16 12:00 AM (7 years, 4 months ago)

99.99, the reason you are seeing bacteria in your spawn is because you are using antibiotic agar... I know it sounds backwards, but antibiotic agar doesn't actually kill bacteria, it just prevents bacteria from flourishing...when you inoculate your jars with a colonized wedge of antibiotic agar the bacteria is still there, you just can't see it...so you are actually just putting it right into the jar...

To prevent this you should transfer from your antiobiotic agar dish on to a dish of regular PDA or MEA agar first, then use that dish to knock up your master jars...

The only real use antibiotic agar has in mycology is to get clean cultures from exceptionally dirty fruits or spore prints....it can actually cause more of a headache than its worth...

Runsfrombears- if you give me more info on your technique and how exactly you are knocking up your masters I may be able to help you out...2


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Offline99.99
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Re: Need help isolating cause of bacterial contamination. [Re: PsilocyBen17]
    #23627685 - 09/09/16 12:26 AM (7 years, 4 months ago)

Quote:

PsilocyBen17 said:
99.99, the reason you are seeing bacteria in your spawn is because you are using antibiotic agar... I know it sounds backwards, but antibiotic agar doesn't actually kill bacteria, it just prevents bacteria from flourishing...when you inoculate your jars with a colonized wedge of antibiotic agar the bacteria is still there, you just can't see it...so you are actually just putting it right into the jar...

To prevent this you should transfer from your antiobiotic agar dish on to a dish of regular PDA or MEA agar first, then use that dish to knock up your master jars...

The only real use antibiotic agar has in mycology is to get clean cultures from exceptionally dirty fruits or spore prints....it can actually cause more of a headache than its worth...

Runsfrombears- if you give me more info on your technique and how exactly you are knocking up your masters I may be able to help you out...2




Yes i know this ... Nomater what i did ore how carful i was around 20% of my bags would contaminate with bacteria
I did this for about four months and said fuck it its not worth the hasel
So even though i still have a problem with bacteria its controled to the point a get three ore four good flushes off my grain in a bag


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Offlinerunsfrombears
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Re: Need help isolating cause of bacterial contamination. [Re: 99.99]
    #23628159 - 09/09/16 06:34 AM (7 years, 4 months ago)

I noc up my spawn jars with mutispore syringe purchased from reputable vendor.
Pc Wbs 60 mins, alcohol wipe lids, heat syringe with lighter, wipe with alcohol wipe, insert syringe thru self healing port.
Like I said. I haven't had my nocs go sour. They look pretty strong actually.


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OfflinePsilocyBen17
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Re: Need help isolating cause of bacterial contamination. [Re: 99.99]
    #23628372 - 09/09/16 08:36 AM (7 years, 4 months ago)

Quote:

99.99 said:
Quote:

PsilocyBen17 said:
99.99, the reason you are seeing bacteria in your spawn is because you are using antibiotic agar... I know it sounds backwards, but antibiotic agar doesn't actually kill bacteria, it just prevents bacteria from flourishing...when you inoculate your jars with a colonized wedge of antibiotic agar the bacteria is still there, you just can't see it...so you are actually just putting it right into the jar...

To prevent this you should transfer from your antiobiotic agar dish on to a dish of regular PDA or MEA agar first, then use that dish to knock up your master jars...

The only real use antibiotic agar has in mycology is to get clean cultures from exceptionally dirty fruits or spore prints....it can actually cause more of a headache than its worth...

Runsfrombears- if you give me more info on your technique and how exactly you are knocking up your masters I may be able to help you out...2




Yes i know this ... Nomater what i did ore how carful i was around 20% of my bags would contaminate with bacteria
I did this for about four months and said fuck it its not worth the hasel
So even though i still have a problem with bacteria its controled to the point a get three ore four good flushes off my grain in a bag




OK but are you transferring the antibiotic stuff to regular agar before inoculating? There should be no reason for a 20% contam rate with the tools you are using. Something is off with your technique. Getting flushes from bacterial bags doesn't sound fun (or safe)


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OfflinePsilocyBen17
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Re: Need help isolating cause of bacterial contamination. [Re: runsfrombears]
    #23628792 - 09/09/16 11:43 AM (7 years, 4 months ago)

Quote:

runsfrombears said:
I noc up my spawn jars with mutispore syringe purchased from reputable vendor.
Pc Wbs 60 mins, alcohol wipe lids, heat syringe with lighter, wipe with alcohol wipe, insert syringe thru self healing port.
Like I said. I haven't had my nocs go sour. They look pretty strong actually.





Ms should really only be used for PF tek style jars. Imo spores>grain is the biggest mistake people make these days as spores are never harvested under sterile conditions. Clean them up using agar first for a higher success rate


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Offlinerunsfrombears
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Re: Need help isolating cause of bacterial contamination. [Re: PsilocyBen17]
    #23629361 - 09/09/16 03:05 PM (7 years, 4 months ago)

Anyone else have any experience with this sort of problem?


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Offline99.99
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Re: Need help isolating cause of bacterial contamination. [Re: runsfrombears]
    #23629864 - 09/09/16 05:26 PM (7 years, 4 months ago)

If i can get growth like this using antibiotic agar, why would i change



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OfflinePsilocyBen17
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Re: Need help isolating cause of bacterial contamination. [Re: 99.99]
    #23630107 - 09/09/16 06:23 PM (7 years, 4 months ago)

I'm done here...


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