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InvisibleJosex
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Re: contamination pic of the day [Re: pixelpopper]
    #24789994 - 11/17/17 05:10 PM (6 years, 2 months ago)

Suppose you vent for 10 min, so the jars get up to temp plus the time it takes to bring the pressure up to 15 psi and then waiting for the pressure to drop before opening the pc... of course mold and spores will be killed dead. Pretty sure they get killed by venting alone.


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InvisibleJosex
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Re: contamination pic of the day [Re: Josex]
    #24797866 - 11/21/17 01:39 PM (6 years, 2 months ago)



Meshed-in mold.

This time it didn't throw a delta as it usually does, but a waxing crescent.


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Invisiblevan hattonFacebookDiscord
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Re: contamination pic of the day [Re: Josex]
    #24797929 - 11/21/17 02:11 PM (6 years, 2 months ago)

Quote:

Josex said:
Does the mold sporulate after a while on the plate or just a couple days after you transferred? Does it grow in little colonies near the edges of the plate? If the mold starts to show and sporulates many days after you transferred, then maybe the filter or lid might be failling.
If it's mold from bad technique it usually shows after 2 or 3 days.
If it's really meshed-in mold, that shit won't sporulate, it is usually some sort of pin mold. I don't think you can get meshed-in trich for instance, that shit will sporulate no matter what, but maybe I'm wrong and you can get meshed-in trich, would like to hear some opinions on this.








I've never heard the term Delta before could someone elaborate?


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If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you


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InvisiblebodhisattaMDiscordReddit
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Re: contamination pic of the day [Re: van hatton]
    #24797945 - 11/21/17 02:15 PM (6 years, 2 months ago)

trich is the most meshed in contamination you can get since it's a parasite of other mycelium.

delta usually means a change or variation.


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InvisibleJosex
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Re: contamination pic of the day [Re: van hatton]
    #24797955 - 11/21/17 02:18 PM (6 years, 2 months ago)

It's like a triangle of uncolonized agar, well, it is colonized but by mold myc growing under the agar. It usually starts to happen when the plate is more than half way colonized.

Also, if there's meshed-in mold myc growth will only be superficial and weak.


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Re: contamination pic of the day [Re: Josex]
    #24797959 - 11/21/17 02:20 PM (6 years, 2 months ago)

Quote:

Also, if there's meshed-in mold myc growth will only be superficial and weak.



not necessarily. look up some of frankhorrigans old posts with hidden contamination. RR was helping him out with that like 3-4 years ago


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InvisibleJosex
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Re: contamination pic of the day [Re: bodhisatta]
    #24797962 - 11/21/17 02:22 PM (6 years, 2 months ago)

Did he get rid of it in the end?


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InvisiblebodhisattaMDiscordReddit
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Re: contamination pic of the day [Re: Josex]
    #24797964 - 11/21/17 02:23 PM (6 years, 2 months ago)

I don't think so. I think it was a start over situation.


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InvisibleJosex
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Re: contamination pic of the day [Re: bodhisatta]
    #24797981 - 11/21/17 02:32 PM (6 years, 2 months ago)

I can't start over though, it was PE from swabs and I have none right now.
I did clean some cultures doing some tricks though, but it's a lengthy process that's not always effective.


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Re: contamination pic of the day [Re: Josex]
    #24797997 - 11/21/17 02:42 PM (6 years, 2 months ago)

that needle poke tissue culture method has got to come in handy. hot pour too

fancy labs would try to incubate cultures at a temperature either so cold or so hot that only one of the organisms survives, hopefully the intended one.


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Invisiblevan hattonFacebookDiscord
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Re: contamination pic of the day [Re: bodhisatta]
    #24798015 - 11/21/17 02:48 PM (6 years, 2 months ago)

I asked and then thought about Delta's in rivers. :lol: but thank you


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If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you


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InvisiblebodhisattaMDiscordReddit
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Re: contamination pic of the day [Re: van hatton]
    #24798135 - 11/21/17 03:38 PM (6 years, 2 months ago)

expensive airline company :cookiemonster:

ha


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OfflineCrispykoot
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Re: contamination pic of the day [Re: van hatton]
    #24799322 - 11/22/17 06:27 AM (6 years, 2 months ago)

Quote:

van hatton said:
I asked and then thought about Delta's in rivers. :lol: but thank you



The contaminant does look like a delta in a river...You will see a point of germination or clustered growth and then it will form a triangle shaped Delta of growth extending from that point that is usually not growing with the mushroom culture.

From observation with meshed in molds I have found that refrigeration of "Clean" looking plates may be a key to seeing the hidden stuff. I see new contaminant growth on plates fresh out of the fridge sometimes... White molds can grow well at low temps. Just look outside..Stuff you would not see otherwise...


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:gd_icon: Shadowboxing the apocalypse and wandering the land :gd_icon:


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InvisibleUllr
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Re: contamination pic of the day [Re: Crispykoot]
    #24803436 - 11/24/17 06:45 AM (6 years, 2 months ago)

I'm getting some bacterial nasties in my agar plates, presumably making their way through the polyethylene film via ridiculous amounts of condensed water.

A couple of days ago I noticed the tiny bacterial dots at the edges of my plates and prepared everything for transfer but, unfortunately, I had some unexpected business to attend and had to drive off after pouring some new plates.

I did the transfer today and contamination had extended a great deal.
Also, my mycelium looks weird, way too fluffy, may that be because of the excess humidity or lack of gas exchange? Polyethylene is supposed to allow gas transfer, but with all the condensation most plates were closed shut, lid stuck to plate by vacuum.

An example of the condensation on a plate where no mycelium grew:



The contaminated plates:



Closeup on the mycelium (I'm guessing it's mycelium, I hope I'm not transferring some weird crap):



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InvisibleTheMadHatter420
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Re: contamination pic of the day [Re: Ullr]
    #24803446 - 11/24/17 06:59 AM (6 years, 2 months ago)

They didn't make it "through" your film. They came from your technique. Either when you poured the plates or made the transfers. Work on your sterile tek. Bacteria doesn't just sneak in like that.

What species are you supposed to be growing on there? Are those germination plates or from transfers and if so how many transfers in are you??


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InvisibleUllr
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Re: contamination pic of the day [Re: TheMadHatter420]
    #24803489 - 11/24/17 07:36 AM (6 years, 2 months ago)

Quote:

TheMadHatter420 said:
They didn't make it "through" your film. They came from your technique. Either when you poured the plates or made the transfers. Work on your sterile tek. Bacteria doesn't just sneak in like that.

What species are you supposed to be growing on there? Are those germination plates or from transfers and if so how many transfers in are you??




Cubensis Z strain, MS from a spore syringe.

If the contam was there from the beginning, how did the bacterial specks appear when mycelium had already taken hold, a week after MS inoculation, and at the very edges of the plate? This is pretty aggressive bacterial growth, from barely visible to photo in around 35 hours.
Other reasons for me to think it comes from around the lid:
The fact that two out of 10 initial plates had a couple of specks of that same growth on day 1, and not at the very edge - meaning it should have shown up earlier on.
The fact that I've noticed very thin bacterial spots at the rim of some plates.
The fact that the jars I inoculated are not presenting signs of bacterial growth (or any growth for the matter, still clean, no clumps, no sticky stuff or wet look).

This is my first grow, and I may be totally wrong, but it looks like that to me. And since water is dropping out through the film, capillarity is totally possible.

Even if my plates look condensation-free after pouring and cooling, they get condensation after taping.
I should add this place is very high-humidity and pretty unsanitary. Even the windows rain down condensation if there are more than two people in the room. So I don't know what to do with that.

If it's my technique, it will be a matter of practise and learning how to work faster. I've been really careful, although too slow being my first time.


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Re: contamination pic of the day [Re: Ullr]
    #24803520 - 11/24/17 07:59 AM (6 years, 2 months ago)

Did you keep any test plates that weren't used to germinate spores? More than likely it is your technique if you just started agar work..practice,practice,practice


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InvisibleTheMadHatter420
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Re: contamination pic of the day [Re: Leftfield420]
    #24803545 - 11/24/17 08:17 AM (6 years, 2 months ago)

How is it the bacteria got through your film, but doesn't any one else's?

If you enlarge this pic, you will see a lot of condensation on the top plate. All mine had extra moisture on them. These plates went into jars like 2 days ago, when they were about 11 days old. I just tossed the test plate yesterday and it was still clean at close to 2 weeks. I say what I am off of experience. If this is your 1st grow there is a better chance that you made a mistake than capillary action pulling bacteria spore into the dish. If it was mold I would buy that possibly as I have seen mold around edges as the spores drift through the air. But only time I see bacteria it is from us, I have never had satellite bacteria unless I let my hand drift over the plate. Just because it didn't grow right away doesn't mean the spore was not there from the start. Hell I have been working on germinating cold weather wood loving spores. Some of the plates sit for 2 weeks with NO germination of ANY kind and then pop up mold, and I am using sealed NO POUR plates for those.



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InvisibleUllr
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Re: contamination pic of the day [Re: Leftfield420]
    #24803602 - 11/24/17 08:51 AM (6 years, 2 months ago)

Quote:

Leftfield420 said:
Did you keep any test plates that weren't used to germinate spores? More than likely it is your technique if you just started agar work..practice,practice,practice




I didn't, but I still have a clean plate that never grew anything... except in-between the two layers of wrap.

Quote:

TheMadHatter420 said:
How is it the bacteria got through your film, but doesn't any one else's?

If you enlarge this pic, you will see a lot of condensation on the top plate. All mine had extra moisture on them. These plates went into jars like 2 days ago, when they were about 11 days old. I just tossed the test plate yesterday and it was still clean at close to 2 weeks. I say what I am off of experience. If this is your 1st grow there is a better chance that you made a mistake than capillary action pulling bacteria spore into the dish. If it was mold I would buy that possibly as I have seen mold around edges as the spores drift through the air. But only time I see bacteria it is from us, I have never had satellite bacteria unless I let my hand drift over the plate. Just because it didn't grow right away doesn't mean the spore was not there from the start. Hell I have been working on germinating cold weather wood loving spores. Some of the plates sit for 2 weeks with NO germination of ANY kind and then pop up mold, and I am using sealed NO POUR plates for those.






Gotcha, thanks.
Still, have you ever had condensation actually drip over the edges of your agar, accumulate inside the wrap, and drip down over the table?
There may be various factors in play. Maybe the water dripping down is helping pre-existing bacteria spread around the edges.
Or maybe it did get inside the film and it was spread by my newbie habit of picking the dishes to check progress way too frequently.

Not questioning your experience, but if there are other possible vectors in play I want to solve those so I can know where and when things failed.


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InvisibleUllr
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Re: contamination pic of the day [Re: Ullr]
    #24817513 - 11/30/17 06:36 PM (6 years, 1 month ago)

Some nasty bacterial cuts from hidden contaminants in the master culture:




Another pretty awesome scalpel contam. This started off as a huge line of bacteria with a tiny mycelium spot in the middle. I can't believe how aggressively this mycelium has fought the contaminant, it concentrated all its energy in running it over with rhizomorphic growth, and only once it had the bacteria in control it started to produce the thinner and wider growth that can be seen now. It's kinda fun to see how these little organisms interact and compete, and I'm glad I didn't toss it straight away and witnessed the fight:



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