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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers]
    #29022408 -

SupaThaRipper said:
i certainly wasnt angry, just firm in my response and im sorry if i came off that way.



Thank you.

SupaThaRipper
>there is a consensus on here that 99% of the time, contamination comes from grain.

Is there? If asked I would be 100 percent sure that the consensus here is "If you've got contam in the tub, you had it on the plate and didn't notice."

That is certainly my take, based on experience, and reading.

I believe is another consensus here is:

"If you have clean grain jars, you will not get contam in your tub, even if exposed to contam."

I've even heard someone say "If you have clean grain jars, you will not get contam in your tub, even if you shit in your tub."

I would not go that far, but it does underscore the idea that a jar of clean grain mycelium is formidably robust and hard to contam.

You are correct that letting jars fully colonize is good practice. I recommend letting them sit 5 days after they look done on the outside and bottom.

I don't think you even lose any time doing that, they grow faster in the tub after that.

One thing I recommend that a lot of people miss is looking at the bottom of the plate. Contam hiding underneath is common.

SupaThaRipper
>you want resources and quotes though? i dont need to pull names in here. this is my argument, not theirs.

I am not saying "prove it, motherfucker"

I am saying "Is there another site like the shroomery that is worth checking out? What's the site?"

I asked you in the post
Quote:
give me some links and I'll look into it....That includes resources outside here. Or I'd like to even know the links for the sites you're talking about.



I'm very into learning and seeing how people do things.

I know there was one other shroom grow forum, was white text on black background and had a hippie style line art on the front page, but it only had a handful of posts a day, and it seems gone now, from my Google search.

SupaThaRipper
>if anyone is having serious contamination issues, trich in particular. i highly suggest them to go through the steps

I think that half the time or more that there is trich, the trich comes from trich-enhanced coir.

EVERYONE: Two brands of coir that do NOT have added trich that are Plantronix Coco Bliss coir, and Nature's Premium brand. One says it does not, and the other I wrote them and asked. Still pasturize with pouring boiling water, breaking it up, and letting it sit covered for a few hours.

Get those if you can, people starting out, Amazon and ebay carry both, at least in the US.

The TC I mentioned with the wacky trich theory, that you asked me who it was, I won't name him but if he wants to jump on I'll discuss it. His theory is this (paraphrased):

"It doesn't matter if you use coir with trich added, because the boiling water kills it all when you pasteurize it."

This is the most wrong thing I've heard on here. (He does good grows despite this because he's skilled and does everything else right.)

I think that is horrible advice for a noob.

Maybe growing shrooms doesn't require to have the same level of clean you'd have in a DNA research lab. (If it did, we wouldn't do it in the same rooms where we live and sleep)

But it should be at least as clean as you'd have in a commercial food processing operation, even a mom and pop one.

Can you imaging a microbrewery or local bread bakery saying "We can can get grain with added mold or without. It's the same price for each. We use the gain with mold because the mold probably all dies when we pour on boiling water."

NO ONE would buy that beer or bread, and they'd be rightfully shut down.

Pic tax, x7x+ F5, 16-quart tub

Edited by nektar61 (11/13/24 05:00 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61] * 1
    #29022442 -

This is too much. I'm done arguing about it lol

Although I do agree with the last half of your post. And I still had issues with coco bliss before Sterilizing

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Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61] * 4
    #29022450 -

Quote:
The TC I mentioned with the wacky trich theory, that you asked me who it was, I won't name him but if he wants to jump on I'll discuss it. His theory is this (paraphrased):

"It doesn't matter if you use coir with trich added, because the boiling water kills it all when you pasteurize it."

This is the most wrong thing I've heard on here. (He does good grows despite this because he's skilled and does everything else right.)

I think that is horrible advice for a noob.



Trichoderma exists everywhere. I highly doubt that your coir contains commercial Trichoderma.  Even if it did, it would be taken care of using boiled water + bucket tek. Your work environment, clothes, hair, and skin is covered in mold spores including those of Trichoderma spp.

Trichoderma infused coir is labeled as such and pretty much exclusively used as seed starter for nurseries. Trichoderma and other beneficial microbes are expensive and are not haphazardly used in the production of coir. Certainly not for coir intended for animal bedding, and definitely not used for coir intended for salt based a/b hydroponics gardening.

The only brand I'm aware of that people often cite is Canna coir.

From Canna:

Quote:
Trichoderma Myth

Trichoderma spp., or any other microbe for that matter, is not inherent in, added to, or cultured for the coco medium CANNA uses. It simply is not there, and, under the strong UV rays of the sun for months, it will not survive as a spore. However, it is one of the most prolific and widely distributed soil microbes in the world. It is in the air from the tropics to the poles, and will not only populate soils, but paper, wood, walls, anything with cellulose. Trichoderma spp. are ubiquitous colonizers of cellulosic materials and can be found wherever decaying plant material is found. Trichoderma will be found in the soil, in the factory, in the home, in the store, anyplace. Some will grow on drywall on the paper covering, on untreated wood doors and trim, anywhere. So the contamination point or source can also be anyplace once the product is exposed.



https://www.canna.ca/articles/story-canna-coco-renewable-growing-medium

They do however carry a seed starter product marketed specifically to nurseries  that does contain Trichoderma.

As stated above, the Trichoderma issue is moot because it is ubiquitous. Use very hot or boiling water to prep CV, no issues. All common moulds and their spores will be destroyed in seconds when exposed to boiled water.



The moment you plunge your hands and forearms into a tub filled with freshly hydrated coir, you're inoculating it with skin bacteria, moulds, yeasts, etc. This is why clean spawn is king. No matter what you do with your substrate, whether it be sterilized, hydrated with boiled water or properly pasteurized, you're mixing environmental contamination throughout when spawning.

Don't grow on substrates previously infested with live moulds including Trichoderma. Process substrates correctly, use clean spawn.




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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap] * 3
    #29022453 -

Lets get back on topic, folks. This is the agar thread. If any of you would like to engage in general cult discussions, well, we have a thread for that.

Double post for emphasis:lol:

:goodluck:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato] * 2
    #29022551 -

Is this logic correct?

A transfer from 1,6% agar to water agar is introducing nutrition to the water agar.
As far as my understanding of the concept goes, by doing this, you have a very clean portion which has access to nutrients, which allows the mycelium to rapidly expand to keep consuming whatever is available on the low-nutrient agar. This search for nutrients will leave most contamination behind and leave you with a bigger portion of clean mycelium.

A stiff agar mix will also retard any bacterial growth, as bacteria usually require a soft media to grow.

So by doing e.g. LME agar > stiff water agar, you give the mycelium solid room to leave any contamination behind, leaving you with a clean and healthy culture.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: MycCycle]
    #29022553 -

This is honestly a very interesting question and I think you should get in depth with playing around with it a bit. I would look forward to seeing what you come up with.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers]
    #29022585 -

SupaThaRipper said:
This is honestly a very interesting question and I think you should get in depth with playing around with it a bit. I would look forward to seeing what you come up with.



I’ll definitely be looking towards shaving off the need for transfers as well as increasing success rate.

An actual experiment with a larger sample size is not going to happen anytime soon, but I’ll be looking to germinate spores on 1,6-2%/2% ELME/Agar, then go straight to sequestering at >2,5% water agar. I’m hoping this will separate mycelium from contamination and suppressing any bacteria susceptible to piggybacking, spores - clean culture with just two plates would save so much time and effort. Not that I don’t enjoy working with plates, but unnecessary work is well, unnecessary.


However, to do this effectively I’d need tons of plates and a microscope + a lot more experience than I currently can brag about.
I was hoping that someone had tried this before. Not necessarily for mycology, but in a general lab setting as well.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: MycCycle] * 1
    #29022590 -

Although interesting for sure, I just find it unneeded in my area, especially if working from spore. I've found that beginning even from the dirtiest of wild prints, using serial dilution to water agar gives your miraculous results, leaving any other methods behind.

I still use a very soft gellan gum. .8% (probably comparable to 1.2% agar?) for spore germination, regardless of bacteria liking softer, without issues

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers]
    #29022618 -

How does water agar compare to brf pucks for cleaning up cultures? I know the brf puck should allow the myc to leave almost any bacteria behind. Water agar serves the same purpose of cleaning bacteria? Or it also helps with enmeshed molds? (Although at this point I don’t even know if enmeshed molds are a real thing or just cultures sectoring)

Nichrome said:
Can someone please show me what "enmeshed mold" is? I have honestly never seen it happen.

That is sectoring. That type of sectoring is indicative of a colony with internal competition going on. Like some of the hyphae are planning a mutiny or like a case of star bellied sneetches vs. regular sneetches. Possibly just a genetic defect. Long story short those patterns often lead to susceptibility to mold taking over but there is no mold in that culture.



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Re: AGAR ENVY! (Anything and All things agar!) [Re: Radamanto] * 3
    #29022635 -

I spent some time playing with water agar, charcoal agar, black tea agar. All slow growth to a crawl. They have their place, but generally I see them more for extreme situations, most agar cleaning can be done with a basic Lme/mea recipe.

That said, I have seen plenty of enmeshed mold, and I hope you and Nichrome never have to.


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Radamanto] * 1
    #29022637 -

My favorite methods in order for dealing with bacteria

Brf puck or agar trench
Agar sabdwich
Water agar

Enmeshed definitely a real thing.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers]
    #29022657 -

I was recently gifted a single dry stipe of new cube genetics, about 4". No cap or gill fragments.

I have tried twice, broke about an inch off, dug out unexposed samples to 4 plates of standard LME.

Each time, the only result is pinkish mold, fusarium? Anyway, the outside must be coated in mold spores.

I have about enough stipe for one more break to unexposed interior. Looking for suggestions, thanks.


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

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Re: AGAR ENVY! (Anything and All things agar!) [Re: LadysKnight]
    #29022665 -

Ahh man I've never had success cloning dry fruits. 😢 Even when very clean they never take off for me.

Gill frag would be amazing. What is the variety?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers]
    #29022667 -

I think I've heard of peroxide soaks? Maybe it was alcohol? Soak the stem in there, should kill any bacteria while maybe allowing for mycelium to still survive. Don't quote me on that


--------------------
I call them Huskies cause they Mush!


The Official Wild Cubensis Thread
Instant Pot Agar
Gnocchi Water Agar
Let's Talk Cowtek

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Re: AGAR ENVY! (Anything and All things agar!) [Re: LadysKnight]
    #29022673 -

In a chance that there could be residual, dry spores, I would take a piece of exterior tissue to 2ml centrifuge filled with polysorbate water and blend on a vortexer. Let sit for 24 hours and then proceed with serial dilution to water agar.

Don't get your hopes up tho

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Huskies]
    #29022675 -

Hydrogen peroxide. But with the lack of good spores, it could be more damaging

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Huskies]
    #29022676 -

Huskies said:
I think I've heard of peroxide soaks? Maybe it was alcohol? Soak the stem in there, should kill any bacteria while maybe allowing for mycelium to still survive. Don't quote me on that



I have seen someone here mention wiping down the stipe with ISO before splitting, but that was specifically for a fresh fruit.


--------------------
"For the powerful freedom-takers who rob justice from us all, that's why I keep my pistol loaded, powder, cap, and ball..."
Read more, post less!
...turns out, I'm an amateur at everything

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers] * 1
    #29022677 -

SupaThaRipper said:
Ahh man I've never had success cloning dry fruits. 😢 Even when very clean they never take off for me.

Gill frag would be amazing. What is the variety?



Variety unknown. But it's a fatty, non-hollow. It would be nice to have, but this is mostly for my own knowledge/experience progression, thanks.


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers] * 1
    #29022679 -

Why the heck would he use serial dilution?
Or polysorbate? Why any of that?

@LN

The odds of tissue revival is close to zero due to the temps reached in the dehydrator. Follow regular cloning practices.

Serial dilution would be total nonsense. Also, vent your pressure cookers, folks.:facepalm3:

https://www.shroomery.org/forums/showflat.php/Number/28789802#28789802

Have you been there yet?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: LadysKnight]
    #29022692 -

I’m only asking/researching about enmeshed molds because my cultures started looking weird. My first batch of plates all looked tomentose with 0 rhizo growth but I could tell they were cubes and even grew decent pf cakes. On this new batch almost all my transfers are looking like this:



The myc used to grow on the surface of the agar but now it’s fuzzier, and growing a bit upwards, more tridimensional growth.

To be fair, I did change my agar recipe a bit, my first batch was 2% nutes 2% agar and the new is 1% nutes 1.5% agar (I was trying new mixes to get some rhizo growth) and also the new plates are cheap chinese ones so they are a bit suspicious.

@LadysKnight does this look like what you described as enmeshed mold?

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