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InvisibleLocN9ne
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Re: contamination pic of the day [Re: PinPornProducer]
    #23046318 - 03/25/16 09:58 PM (7 years, 10 months ago)

Damn, that's discouraging as fuck... Definetely try upping the time...


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OfflinePinPornProducer
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Re: contamination pic of the day [Re: LocN9ne]
    #23046367 - 03/25/16 10:14 PM (7 years, 10 months ago)

Yea it almost def has to be a time issue


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InvisiblePastywhyteMDiscord
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Re: contamination pic of the day [Re: PinPornProducer]
    #23046443 - 03/25/16 10:30 PM (7 years, 10 months ago)

Mold is 99% of the time in cases like yours a filter fail. Liquid media, like agar reach mold killing temps in 10 min. I have only seen bacteria from insufficient cycle times. If it was only in one plate that really points to filter fail.


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OfflinePinPornProducer
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Re: contamination pic of the day [Re: Pastywhyte]
    #23046513 - 03/25/16 10:50 PM (7 years, 10 months ago)

I tossed all remaining 8, I wasn't going to risk using any of them. I'll look into the filter...


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OfflineShroomsNB
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Re: contamination pic of the day [Re: Pastywhyte]
    #23046521 - 03/25/16 10:51 PM (7 years, 10 months ago)


I got some green in my grain that suked and mono ruined my mood


Edited by ShroomsNB (03/25/16 10:59 PM)


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Offlinetwistedty
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Re: contamination pic of the day [Re: PinPornProducer]
    #23046615 - 03/25/16 11:18 PM (7 years, 10 months ago)

Quote:

PinPornProducer said:
So the 21st of last month I made some pasty plates, I only needed 4 but made 12 and stored the other 8 in ziplocs and kept them in my gb. Go to use a few of them today and seen this in 1 of the plates...



None of the filter was touched, I never even removed the paper towel pieces from the lid.



Pc longer? The last few batches were only pcd for 30 min which I feel was plenty long enough but maybe I should go back to 45 min :shrug:




i do 45. and do my work in a SAB


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Invisibleblindingleaf
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Re: contamination pic of the day [Re: twistedty]
    #23046622 - 03/25/16 11:20 PM (7 years, 10 months ago)

thats filter/silicone failure


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MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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Offlinetwistedty
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Re: contamination pic of the day [Re: blindingleaf]
    #23046697 - 03/25/16 11:39 PM (7 years, 10 months ago)

Quote:

blindingleaf said:
thats filter/silicone failure





+5 on that sig pic.

damn


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InvisiblePastywhyteMDiscord
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Re: contamination pic of the day [Re: twistedty]
    #23047278 - 03/26/16 07:07 AM (7 years, 10 months ago)

Quote:

twistedty said:
Quote:

blindingleaf said:
thats filter/silicone failure





+5 on that sig pic.

damn




True Dat.  leaf makes boss hog look like a piglet :rofl:

:likeaboss:


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Invisibleblindingleaf
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Re: contamination pic of the day [Re: Pastywhyte]
    #23047291 - 03/26/16 07:12 AM (7 years, 10 months ago)

only when my airstone and heating pads are working correctly though :crazy:


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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InvisiblePastywhyteMDiscord
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Re: contamination pic of the day [Re: blindingleaf]
    #23047297 - 03/26/16 07:15 AM (7 years, 10 months ago)

Quote:

blindingleaf said:
only when my airstone and heating pads are working correctly though :crazy:




:lolsy:


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InvisibleSupalemonhaze
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Re: contamination pic of the day [Re: Pastywhyte]
    #23048002 - 03/26/16 12:31 PM (7 years, 10 months ago)

Agar should be sterile after 10 mins at 15psi. I make batches of 600ml in glass bottles pced for 15mins and never ran into problems. When I used pasties I gave them the same pc cycle and all went well.

It was only 1 plate that contaminated right? My best guess is that it happened while it was cooling, sucking in air. This happened to me a couple times. What filter are you using? If it's something like micropore tape just change it but don't read too much into it.


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Offlinetwistedty
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Re: contamination pic of the day [Re: Supalemonhaze]
    #23048250 - 03/26/16 02:00 PM (7 years, 10 months ago)

Quote:

Supalemonhaze said:
Agar should be sterile after 10 mins at 15psi. I make batches of 600ml in glass bottles pced for 15mins and never ran into problems. When I used pasties I gave them the same pc cycle and all went well.

It was only 1 plate that contaminated right? My best guess is that it happened while it was cooling, sucking in air. This happened to me a couple times. What filter are you using? If it's something like micropore tape just change it but don't read too much into it.





that is a very short cycle.  i dont see any reason to go that short.


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InvisiblePastywhyteMDiscord
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Re: contamination pic of the day [Re: twistedty]
    #23049574 - 03/26/16 09:02 PM (7 years, 9 months ago)

10 min might be barely acceptible for lab grade agar and nutrition but I would never trust a run that short for the solids etc that we use with many home based recipes. Using expensive MEA so I can short my cycle isn't worth the upside to media rotation and culture preference.


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InvisibleSupalemonhaze
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Re: contamination pic of the day [Re: Pastywhyte]
    #23054080 - 03/28/16 08:22 AM (7 years, 9 months ago)

15mins isn't that short IME. I mentioned 10 minutes because they were talking about it a few months back and someone mentioned that media such as agar and LCs are usually sterile after 10 mins.

I started PCing for 20-25 mins when I started due to begginers paranoia but once I started reading all these different teks I noticed there's a bunch of others PCing for 15 mins. Gave it a go and never got contams in my blank plates.

Quote:

Pastywhyte said:
Using expensive MEA so I can short my cycle isn't worth the upside to media rotation and culture preference.




All malt extract is expensive here, so I don't know what consistency cheap ones are. I use brewers malt so I guess that is good quality.

What do you mean by culture preference? I use GWA sometimes to mix it up but I haven't got to the point where I actually needed to change my media yet. I prefer malt since I have to defrost my grain water to be able to use it so that's what I use for the most part.

Even with GWA though, I use the same cycle without any headaches. I also PC glass petris at the same time I PC my agar and they don't cause me any trouble.

How long do you guys PC your agar for then?

At any rate 30mins is more than enough time to sterilise agar I reckon, I doubt that was the problem in pinporn's instance.

Edit:

In reality when I say 15 mins at 15psi that's not counting the time it takes to get to 15 and the time it takes to drop to zero. With agar I don't remove the weight like I do with grain jars since it's easy to boil it over so it's under high heat for at least 7-10 extra minutes.


Edited by Supalemonhaze (03/28/16 08:28 AM)


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InvisiblePastywhyteMDiscord
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Re: contamination pic of the day [Re: Supalemonhaze]
    #23054106 - 03/28/16 08:36 AM (7 years, 9 months ago)

How many runs have you done with anything other than malt for 10 min? I have had bacillus contam every plate with a run as short as 25 min. Yes 9 times out of 10 with a filtered broth and low population media you can get away with it. But that 10th time when it fails you will be kicking yourself for skimping those minutes.

Malt is pricy compared to other cheaper items. Potato flakes are excellent for cubes but need longer due to the solids they contain. At any rate I believe in media rotation so just using malt is a poor option regardless of how short you can run it.

If it works then it works. But I have been bitten a few times by skimping out on something as simple as just leaving the heat on an extra few min. So I recommend a safe cycle, that is a foolish corner to cut IMO. You might agree with me if it ever happens to you. Only takes once to ruin a lot of projects.


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Re: contamination pic of the day [Re: Pastywhyte]
    #23054133 - 03/28/16 08:50 AM (7 years, 9 months ago)

how has this thread gone under my radar for 6 fuckin months?!


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Invisibleblindingleaf
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Re: contamination pic of the day [Re: Munchauzen]
    #23054341 - 03/28/16 10:06 AM (7 years, 9 months ago)

I do 30-40 minutes for two bottles awith 500ml each for all recipes.  The only time I've read about reducing cycle time is with gentamincyn so its not "deactivated" but the few times I used it, 30 min seemed to work fine

The only times I've seen bacteria on uninnoculated agar is when i used ground WBS or rolled oats, and it took about a month at room temp to show, always on the bottom where the solids landed during gelling


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A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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OfflineGreenRabbit
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Re: contamination pic of the day [Re: blindingleaf]
    #23054737 - 03/28/16 12:04 PM (7 years, 9 months ago)

Whoa, how did thread slide past me for 6 months?

I'll add some new pics soon.

For now, here's some old ones of what happens to a PF jar if not sterilized (not inoculated either).
I can see at least 5 different contaminants.



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InvisibleSupalemonhaze
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Re: contamination pic of the day [Re: Pastywhyte]
    #23055672 - 03/28/16 04:40 PM (7 years, 9 months ago)

Quote:

Pastywhyte said:
How many runs have you done with anything other than malt for 10 min? I have had bacillus contam every plate with a run as short as 25 min. Yes 9 times out of 10 with a filtered broth and low population media you can get away with it. But that 10th time when it fails you will be kicking yourself for skimping those minutes.

Malt is pricy compared to other cheaper items. Potato flakes are excellent for cubes but need longer due to the solids they contain. At any rate I believe in media rotation so just using malt is a poor option regardless of how short you can run it.

If it works then it works. But I have been bitten a few times by skimping out on something as simple as just leaving the heat on an extra few min. So I recommend a safe cycle, that is a foolish corner to cut IMO. You might agree with me if it ever happens to you. Only takes once to ruin a lot of projects.




I've used GWA about 3 or 4 times so far, I always do 15mins though not 10. But I pass it through coffee filters so there isn't any solids, probably why I never had problems.

Never knew I was actually cutting a corner, was under the idea that 15 mins is plenty for water based stuff. Never tried PDA. I will keep this in mind if I ever need to use medias other than MEA or GWA. How long do you think until cultures need their media to be changed? I was under the impression that that takes months at least. Also changing between MEA and GWA should be enough of a change for the mycelium to recover right?


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