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Jars of "Fail" for us Newbs


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#1 Jinroh

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Posted 21 November 2019 - 03:02 AM

Hello, fellow Newbs!!

 

When I first started this hobby six months ago, one of my major questions was how do I know that I am looking at Mycelium or something bad like mold or other invasive fungi? Since then I see this question almost daily (mainly in the new member introduction thread).

 

I thought I would post some images of contaminated spawn jars and an example of a healthy growing jar and a fully colonized jar.

 

I am not claiming anything in this post, I am not an expert on mold and fungi identification. I am just posting this because I feel it might be of some use to people like myself just getting our feet wet (so to speak).

 

What I AM an expert at (as the photos will show) is causing and growing contamination. I recommend NOT getting good at that skill as I have.

However, it is sure to happen. The photos of contamination are all from the same inoculation day and exhibit at least three types of contamination... maybe more! I purposefully inoculated 25% more jars than I need because I expect contamination. Maybe when I get a flow hood and more experience I will not have to waste so much spawn, but until that day comes I feel making more spawn than needed is a good tactic for a beginner and I recommend it.

 

As I said, the below images are of the same inoculation day, from the same SAB and the same grain from the same pressure cooker (just several batches). All the jars were stored in my fridge for about two weeks before inoculation. Beginners will find that inoculation day involves a fair amount of prep so doing it all at once is advisable. IMHO

For each contaminated jar you see, there are at least two successfully colonized jars that have since been put into a sub.

 

IMG_0923.jpg IMG_0924.jpg IMG_0925.jpg IMG_0926.jpg IMG_0927.jpg IMG_0928.jpg

 

Below is early stage colonization from Agar that is around 7 days old. And another of a fully colonized jar next to a rancid jar for comparison. The good jar is a snowy white in contrast to the contaminated one. What is important to note, the contaminated jars looked perfect up until about a week ago. I learned because of that, to wait to see pins form in the jar before birthing. That way you reduce the chance of transferring bad spawn into good spawn and ruining it all. That is not to say it still won't happen but you will be off to a better start!

Also very important! NEVER open a contaminated tub or jar in your house! You will most likely ruin any future grows by airborne contamination. I take my jars outside far from the house, wear a dust mask, and slowly fill them with a 50/50 mix of bleach and water and let them sit 30 minutes and then throw the spawn in the trash and resanitize the jars and lids.

 

        Good verses bad                                         Early Stage Colonization from Agar to fully colonized                 

IMG_0937.jpg                                      No contamination visible yet                           

                                                                                                 good and done.jpg      

Same bad jar above early stage contamination

earlystage.jpg

 

Pinning jar ready to birth

IMG_0936.jpg                                                                                                                 


Edited by Jinroh, 21 November 2019 - 03:27 AM.

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#2 RutgerHauer

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Posted 21 November 2019 - 04:32 AM

How far away from your house do you go when getting rid of it? Seems to be unnecessary to go very far.

If you are an expert at contamination, you would be able to avoid it most of the time. :) But I see your point. I feel for you.

Seems to me that the problem is somewhere in the source you use for inoculation, sterilization or SAB technique - or a combination.

Some people rather throw out their badly contaminated jars completely, because fully cleaning and sterilizing them is a real pain. Might have to run your PC for quite a bit longer to be safe.

So make entirely sure your jars are completely sterile, the grain is prepared and sterilized properly (used to run too short a PC cycle at first myself, and had the wrong source of rye grain), your agar plates are clean and your SAB technique is correct. I found that if you don't work in a patient manner things tend to go wrong, once you are cutting corners.

Be methodical, precise and make notes (so you can see where you diverged from your usual method and how that impacts the result)

I am for one also very reluctant to inoculate directly from agar without flow hood.

One solution might be to inoculate an LC jar from agar first to expand your culture, which you can then test before using it to inoculate grain spawn. I have good experience inoculating with syringes, seems to be the safest way to go about it if you only have an SAB.
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#3 Jinroh

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Posted 21 November 2019 - 04:52 AM

How far away from your house do you go when getting rid of it? Seems to be unnecessary to go very far.

If you are an expert at contamination, you would be able to avoid it most of the time. :) But I see your point. I feel for you.

Seems to me that the problem is somewhere in the source you use for inoculation, sterilization or SAB technique - or a combination.

Some people rather throw out their badly contaminated jars completely, because fully cleaning and sterilizing them is a real pain. Might have to run your PC for quite a bit longer to be safe.

So make entirely sure your jars are completely sterile, the grain is prepared and sterilized properly (used to run too short a PC cycle at first myself, and had the wrong source of rye grain), your agar plates are clean and your SAB technique is correct. I found that if you don't work in a patient manner things tend to go wrong, once you are cutting corners.

Be methodical, precise and make notes (so you can see where you diverged from your usual method and how that impacts the result)

I am for one also very reluctant to inoculate directly from agar without flow hood.

One solution might be to inoculate an LC jar from agar first to expand your culture, which you can then test before using it to inoculate grain spawn. I have good experience inoculating with syringes, seems to be the safest way to go about it if you only have an SAB.

I said I am an expert at CAUSING contamination. Not an expert at AVOIDING it. LOL

 

I take the jars at least 200 feet away just to ensure no airborne particles that might escape when I open the jars land on my front porch. I reuse my lids because they are all custom made autoclavable with .2 micron permanent filters and self-healing injection ports.

 

I usually PC my grain and sub for at least 90 (usually more like 100) minutes. My LC liquid for 20 minutes to avoid caramelizing the nutrient.

All of my grain is inoculated from agar plates created from my own homegrown liquid cultures. I test those cultures on three individual agar plates and compare them after colonization to see if I have contamination in all three. If I don't then I am pretty sure my culture is good and I use the best plate to inoculate my jars.

During colonization, I prepare more jars only half full. Once the full jars are colonized I split each full jar into all of the half-full jars effectively doubling my yield. So far the contamination rate is between 25 and 30%. To me, this is acceptable since I live deep in the woods of Appalachia and it is very humid and hot in the warm months and I have a lot of problems with mold and fungus on my home exterior much less in my grain jars. So I know I have contamination in my air. It is unavoidable where I live. So my inoculation room has a very large HEPA purifier that I run for several hours before I even open the SAB. During that time I also spray and wipe down the area, all the jars... everything as is the norm.
The only thing I think I am missing is a flow hood. I am slowly gathering the parts as my budget allows. Anything over a 16-inch grid gets pricey so I am saving my money. I don't want a tiny one. I would like at least 24 inches so if I decide to start selling the "legal" kind at the farmers market, it will be more efficient for me.


Edited by Jinroh, 21 November 2019 - 05:03 AM.

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#4 FunG

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Posted 21 November 2019 - 04:59 AM

Dont store uncolonized sterile grains in the fridge, a fridge is filthy with bacteria.

Its not a problem if the nutrient source is fully run with mycelium, then by all means store in a fridge, but not sterile grow medians and then expect them to still be sterile.

Keep them in a cuboard with the seal intact that forms after pressure cooking and they'll be just fine for a good while.

You know, it's sad someone had to make a thread about this but I totally agree, I've known people who have thrown out perfectly healthy mycelium thinking it was mold..... it's as if they had never seen a pic of a fully colonized jar or something absolutely baffling...

#5 Jinroh

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Posted 21 November 2019 - 05:09 AM

Dont store uncolonized sterile grains in the fridge, a fridge is filthy with bacteria.

Its not a problem if the nutrient source is fully run with mycelium, then by all means store in a fridge, but not sterile grow medians and then expect them to still be sterile.

Keep them in a cuboard with the seal intact that forms after pressure cooking and they'll be just fine for a good while.

You know, it's sad someone had to make a thread about this but I totally agree, I've known people who have thrown out perfectly healthy mycelium thinking it was mold..... it's as if they had never seen a pic of a fully colonized jar or something absolutely baffling...

That is why I started this topic. I knew most people would not need the info but 6 months ago I never had seen a colonized ANYTHING and had to ask questions and share photos and somethimes it is hard to see if it is contam or not. I realize I am not covering every scenario, I am just sharing my limited experience.

 

And as far as storing in my fridge, my lids all have permanent filters with high temp silicone gaskets... I asssumed as long as I sprayed down the jars before inoculation I would be OK.


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#6 RutgerHauer

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Posted 21 November 2019 - 06:39 AM

I understand, dont feel like I'm attacking or anything.. My point was if you were an expert at causing contamination it would also be so that you would be avoiding them successfully for the most part. I think it's a semantic question of what an expert is. Any idiot can contaminate a grow and be very good at it, not everyone can work around those issues like an expert would.

Anyway I see in your response that you are on the right track in trying to limit the problems.

I am unsure about using hepa filters for SAB work is, since the concept is about the air that is still, not clean, and you want as less air movement as possible for a while before starting your work.

The risk in using agar for inoculation with SAB is, I think, not that your plate isn't clean, but as soon as you open up a jar to inoculate, you risk contaminants getting in there a lot more than for example using a syringe/SHIP. That's why I propose to work from agar to LC to grain and not from LC to agar to grain. Or at least use your LC directly to grain and cut out the agar step except for using it to test.

I sterilize my grains for 105 minutes now - and the pressure will keep up after that for another 10 to 15 minutes, 90 might not be sufficient, depending on what PC you use.

I also started to sterilize my metal and glass equipment for 30 minutes, though people seem to say 15 is enough.

I have had trouble when doing G2G, where jars seemed clean but contaminated after transferring. This can be due to giving the contaminants extra time and clean grain to take over, but also yet another moment where you expose the stuff to open air for a second time.


Also, I'm no expert at anything either so don't think I think I know it all. :)

Edited by RutgerHauer, 21 November 2019 - 07:17 AM.

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#7 Jinroh

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Posted 21 November 2019 - 06:55 AM

I understand, dont feel like I'm attacking or anything.. My point was if you were an expert at causing contamination it would also be so that you would be avoiding them successfully for the most part. I think it's a semantic question of what an expert is. Any idiot can contaminate a grow and be very good at it, not everyone can work around those issues like an expert would.

Anyway I see in your response that you are on the right track in trying to limit the problems.

I am unsure about using hepa filters for SAB work is, since the concept is about the air that is still, not clean, and you want as less air movement as possible for a while before starting your work.

The risk in using agar for inoculation with SAB is, I think, not that your plate isn't clean, but as soon as you open up a jar to inoculate, you risk contaminants getting in there a lot more than for example using a syringe/SHIP. That's why I propose to work from agar to LC to grain and not from LC to agar to grain.

I sterilize my grains for 105 minutes now - and the pressure will keep up after that for another 10 to 15 minutes, 90 might not be sufficient, depending on what PC you use.

I also started to sterilize my metal and glass equipment for 30 minutes, though people seem to say 15 is enough.


Also, I'm no expert at anything either so don't think I think I know it all. :)

No No! I do not take constructive advice or criticism as an attack. I was merely explaining my current process.

 

Obviously, I have an issue with my process.
I also understand that a flow hood is not the end-all answer to contamination if there is a failure in the process chain leading up to the flow hood step.
A flow hood or a SAB won't do anything if there is a bad hygiene technique. There could very well be a weakness in my process. If there weren't I would have zero contamination.

I do my best to emulate what I see other successful growers do... but that does not mean I actually get it right. That is my whole purpose on this site. To learn and give my personal opinions based on my experience. I always make sure everyone knows that I am still learning and to not take what I say as the right advice. There is a lot of conflicting opinions in this hobby and some people can really get bent out of shape if they feel someone is throwing bad info into the mix. In fact, this is mentioned in the site rules.


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#8 RutgerHauer

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Posted 21 November 2019 - 07:43 AM

I am even more 'expert' at contaminating jars by the way, I recently inoculated a jar of rye with a pure culture of a Trichoderma variety without even realizing. Was trying to cultivate an inactive species that was unknown to me so was unsure what its mycelium is supposed to look like. Some of them look very different. Talking about noob fails...

Of course I threw it out completely since it's useless. But, I took a picture of it that might be relevant here, if people were to wonder what a jar full of trich can look like. (Before speculating and turning green)
Definitely something you won't see on these forums very often voluntarily shared..

You can hardly see the mycelium since it's so fine and wispy. A very big difference compared to healthy thick mycelial growth of Psilocybe Cubensis.

Attached Thumbnails

  • PSX_20191121_133356.jpg

Edited by RutgerHauer, 21 November 2019 - 07:47 AM.

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#9 Jinroh

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Posted 21 November 2019 - 07:50 AM

I am even more 'expert' at contaminating jars by the way, I recently inoculated a jar of rye with a pure culture of a Trichoderma variety without even realizing. Was trying to cultivate an inactive species that was unknown to me so was unsure what its mycelium is supposed to look like. Taking about noob fails...

Of course I threw it out completely since it's useless. But, I took a picture of it that might be relevant here, if people were to wonder what a jar full of trich looks like. (Before speculating and turning green)
Definitely something you won't see on these forums very often voluntarily shared..

You can hardly see the mycelium since it's so fine and wispy. A very big difference compared to healthy thick mycelial growth of Psilocybe Cubensis.

That is a good shot... it looks like a strain of spider mold to me... grayish instead of white which will turn green eventually as it invites more to the party.
I think the more photos that members upload of different contaminations the better it will be for newbs. Hopefully, more people do.
I think some people are embarrassed when in fact it even happens to the pros sometimes... in small and large scale operations.
To me contamination is not a failure... it is a part of the hobby. A goal to shoot for conquering.


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#10 RutgerHauer

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Posted 21 November 2019 - 08:00 AM

I agree. Thought the first instinctual feeling is to be embarrassed, now I have experienced this it is much more likely that I will recognize it and get rid of it sooner. What else I found remarkable about this contaminant is that it smelled like coconut in a later stage on agar. Which lead me to the conclusion it was indeed a trichoderma species.

Here are the agar shots:


Edit: smell is something that can be very useful to determinate, like in this case. Bacterial contaminants often literally smell like shit.

Attached Thumbnails

  • PSX_20191031_233501.jpg
  • 20191110_222433.jpg

Edited by RutgerHauer, 21 November 2019 - 08:05 AM.

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#11 Jinroh

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Posted 21 November 2019 - 08:10 AM

I agree. Thought the first instinctual feeling is to be embarrassed, now I have experienced this it is much more likely that I will recognize it and get rid of it sooner. What else I found remarkable about this contaminant is that it smelled like coconut in a later stage on agar. Which lead me to the conclusion it was indeed a trichoderma species.

Here are the agar shots:

That top left photo looks really freaky. Like a sea anemone or coral.

 

Smell is a good indicator... mine smelled sour.

 

Here is an interesting shot of a plate magnified 400X that I took about two months ago. I took the shot with my phone shooting the screen of my microscope so it is a little blurry. You can clearly see the mycelium fighting the trich and attempting to overrun it. Really cool!

 

 

 

APE Culture Test.jpg micro battle.jpg


Edited by Jinroh, 21 November 2019 - 08:16 AM.

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#12 RutgerHauer

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Posted 21 November 2019 - 08:20 AM

Yeah I thought that bacterial coral pattern was pretty cool looking. That grew out in 24 hours and grew in 'stacks' after that.

Nice close up shot man! You can also see the mycelium fighting spores that landed on top of it on the normal pic, in the form of those droplets.

#13 Jinroh

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Posted 21 November 2019 - 08:21 AM

You know... I was wondering what that droplet was. I assumed it was digestive enzymes.



#14 RutgerHauer

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Posted 21 November 2019 - 08:43 AM

I must say that I am not entirely sure, but I recognize it from a plate of mine that eventually proved contaminated, and it seems to me to be very logical to draw this conclusion.
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#15 buzzkilluton

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Posted 21 November 2019 - 11:55 AM

One thing I have done to lower my contamination rate is when I am running my PC (23qt Presto) I let it vent steam for 10 minutes after the little pressure latch locks in place. This ensures that there is not a pocket of air in your PC and that you are reaching proper temperature and PSI. The last few PC runs have not contaminated at all. Also make sure you are properly preparing your SAB for your work. I used to have a 50% fail rate now I it's more like 1 jar out of 20. Hope this helps man. Keep on trucking man.
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#16 macgyver

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Posted 21 November 2019 - 01:31 PM

Perfect thread to put these pictures my friend sent me!

" How do my PF jars look?"

Snapchat-688715876.jpg Snapchat-1210263469.jpg Screenshot_20191120-080520_Snapchat.jpg


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#17 RutgerHauer

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Posted 21 November 2019 - 01:46 PM

Haha fuck me. THAT is nasty.

Looks like peanuts in wasabi with some chili peppers.

Edited by RutgerHauer, 21 November 2019 - 01:47 PM.


#18 macgyver

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Posted 21 November 2019 - 01:56 PM

Crazyyyy he used a LC that was almost identical to a clean looking one but was red instead. I'm thinking he literally germinated trich and some form of lipstick with a dash of cobweb for good measure. Those jars are not even 7 days after inoculation. Prepped correctly so he just has a culture of every cultivators worst nightmares :biggrin:


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#19 RutgerHauer

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Posted 21 November 2019 - 02:08 PM

I wonder if there is any cubensis in there at all.. I think it will get you fucked up if you eat it though.

#20 Jinroh

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Posted 21 November 2019 - 02:32 PM

One thing I have done to lower my contamination rate is when I am running my PC (23qt Presto) I let it vent steam for 10 minutes after the little pressure latch locks in place. This ensures that there is not a pocket of air in your PC and that you are reaching proper temperature and PSI. The last few PC runs have not contaminated at all. Also make sure you are properly preparing your SAB for your work. I used to have a 50% fail rate now I it's more like 1 jar out of 20. Hope this helps man. Keep on trucking man.

Yep! I know the 10 minute air purge trick from my experience in canning from my garden. I don't start my timer until after 10 minutes of purging and then it has to reach 15 psi before I start it.






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