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Offlineborderline
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second contaminated batch - looking for guidance
    #27120295 - 01/01/21 12:16 PM (3 years, 27 days ago)

Hello all. I recently decided to try to grow and I am enjoying the whole process but running into some hurdles. Hoping for some help on identifying where I can improve my aseptic technique and hopefully identify where I am potentially getting contamination. I'm following BRF tek procedure.

I ordered some Golden Teacher spores from sporestore a little over a month ago. I inoculated 6 jars and all 6 grew a purple mold (fusarium is my best guess, maybe Neurospora?) around 6-7 days post inoc. I deserved that though because I just bought BRF jars off Amazon and I did not re-sterilize (I know, dumb. my excuse was that I wanted to inoculate before leaving for a long weekend and did not have a pressure cooker, rolled the dice based off many in the comments section having success without re-sterilizing)


I properly disposed of all jar contents, sprayed heavily with isopropyl, dish washer, sprayed again and used for another round.

This time I made my own cakes (I'm following Zirian's updated pf tek guide, but also I have read several other guides and watched youtube videos). However, I did stray in that I did not use a pressure cooker and instead used an instant pot (did 3 hours sterilization to make sure). I may just go buy a pressure cooker today.

I ordered 4 different syringes, again from sporestore. I inoculated 9 jars with 3 different PC strains, each strain in 3 jars. At day ~4-5 the 3 jars with "PF classic" started to show signs of mycelium growth, but looked a bit "cottony". ~ day 5-7 they have all started to show a blue/green coloring. From what I understand, this shouldn't be bruising during this stage, and my best guess is it is Penicillium.




The other 6 jars (3 with Hanoi and 3 with Malaysian strain) are just showing small spots indicating mycelium growth today on day 7. Hopefully they are all good.

Feeling a bit demoralized and hoping for some advice on where I can improve. However, this latest round of contam seems like it must be the spores. I did notice some clumping in one of the syringes, but I am unsure which one it was (I read this can be a sign of syringe contamination?) Sporestore seems reputable though, so it certainly could be something I am doing wrong. Should I attach gloves to my SAB to get it more air tight? Should I build a flow hood? I am just doing this as a hobby at home, but I can see myself getting more into it, so I plan on springing for a good pressure cooker as soon as I can justify it. Any guidance is appreciated. :smile:


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OfflineMycoBrainz
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Re: second contaminated batch - looking for guidance [Re: borderline]
    #27120345 - 01/01/21 12:39 PM (3 years, 27 days ago)

Get a pressure cooker and follow a tek to the T. Thats not bruising, that's tric. I just had some problems. Had a friend come watch and we found out my pressure cooking temp was too low. Turned up stove a little and extended time by 20 min, back in sterilization.


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OfflineKyngwhatt
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Re: second contaminated batch - looking for guidance [Re: MycoBrainz] * 1
    #27120367 - 01/01/21 12:52 PM (3 years, 27 days ago)

i personally think the pressure cooker is non negotiable in this hobby, but some people think that it's not necessary for cakes. I know that the people that use a pot try to get a pot with a tight fitting lid that they can weigh down to try and mimic a pressure cooker. if you get that figured out i'd check out my sterile technique because although it does happen but trusted vendors usually put out really good syringes.


Don't be demoralized friend

don't be afraid to ask any questions if you are having trouble finding information.


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Whatt


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Offlineborderline
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Re: second contaminated batch - looking for guidance [Re: Kyngwhatt]
    #27120380 - 01/01/21 01:00 PM (3 years, 27 days ago)

Gonna do another round of inoculation this week. I will be sure to use a pressure cooker :smile:


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Offlineborderline
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Re: second contaminated batch - looking for guidance [Re: borderline]
    #27120385 - 01/01/21 01:03 PM (3 years, 27 days ago)

If it was my sterilization technique, should I expect the other 6 jars to grow tric as well? I would assume so.


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OfflineKyngwhatt
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Re: second contaminated batch - looking for guidance [Re: borderline]
    #27120401 - 01/01/21 01:15 PM (3 years, 27 days ago)

probably going to get the other 6 because it looks like trich took over before the spores even started germinating. hope not but probably :shrug:


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Whatt


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Offline1kldude
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Re: second contaminated batch - looking for guidance [Re: Kyngwhatt]
    #27120415 - 01/01/21 01:21 PM (3 years, 27 days ago)

Looks like cobweb mold along with green mold.

Cobweb mold has a grayish tint and spreads really fast.

I am really good at growing cobweb mold so yeah :laugh:.

When i was a noob which i still am i didnt use a pressure cooker and was soooo amazed that they colonized so quickly to learn it was really just mold :laugh:.


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<<<<----Any post I make is purely fictional and for informational purposes only---->>>>


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OfflineKyngwhatt
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Re: second contaminated batch - looking for guidance [Re: Kyngwhatt]
    #27120428 - 01/01/21 01:27 PM (3 years, 27 days ago)

Just thinking ahead if you have problems next time after using pressure cooker. i'd start looking at my syringes considering that they were plunged into contaminated substrate. the flame sterilizing should have taken care of that its just something i'd want to be aware of.


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Whatt


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Offlineborderline
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Re: second contaminated batch - looking for guidance [Re: Kyngwhatt]
    #27121955 - 01/02/21 12:16 PM (3 years, 26 days ago)

I did flame sterilize between inocs, but it's possible. None of the other jars are showing contam yet


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Offlineborderline
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Re: second contaminated batch - looking for guidance [Re: borderline]
    #27121963 - 01/02/21 12:17 PM (3 years, 26 days ago)

also, the spores came in 4 different syringes/needles.


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OfflineSchemenhaft
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Re: second contaminated batch - looking for guidance [Re: borderline]
    #27122033 - 01/02/21 12:45 PM (3 years, 26 days ago)

Quote:

borderline said:
also, the spores came in 4 different syringes/needles.




You need to go for Agar-plates.
Everything else is just hoping for being lucky.
You get a syringe which is filled with spores of your preferred mushroom - But also other contamination like mold spores and bacteria.
Of course, sometimes you could get a good one which is almost contamination-free and you are lucky your wanted mushroom is faster and strong enough.

But this is always just playing with fire even if you decide to grow on something else then brf.
I did the same way like you, started without PC, without Agar-plates directly from spores to spawn - Forget it, I wasted months of time for like 5g of Shroom and tons of contamination in my house.


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Offlineborderline
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Re: second contaminated batch - looking for guidance *DELETED* [Re: Schemenhaft]
    #27124607 - 01/03/21 07:33 PM (3 years, 25 days ago)

Post deleted by borderline

Reason for deletion: mispelling


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Offlineborderline
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Re: second contaminated batch - looking for guidance [Re: borderline]
    #27159693 - 01/20/21 02:20 PM (3 years, 8 days ago)

update:
No other contam, but some jars are taking while and some appear to have no growth. I need to be more liberal with the liquid culture I guess. I did another round of cake innoc on 1/4 but I think the moisture content is a little off because they are going slower than I would expect. I flipped them a few days back and it seemed to help a few of the jars. I'm considering pulling off any micropore tape to help with gas exchange.

I believe the contam was a bad syringe, no other jars have shown contamination. I think the instant pot works fine, but is not ideal obviously. I have a PC now and I am going to do grain and bulk monotub on the next round. Will post some pics later :smile:


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