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Invisiblebbox244
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First trich contam, quick question on trich
    #14727360 - 07/06/11 05:29 PM (12 years, 7 months ago)

If a WBS master jar is used to transfer colonized WBS to quart WBS jars via g2g and trich spores are introduced into the quart jars, how long will it take the spores to germinate and produce green speculating mold in the quart jars? 

I tried my first g2g (I normally only do GLC's) and this was to whip up a batch of tincture, and the 2 g2g based trays turned green under the surface before full colonization after 18 days, while the 4 GLC based trays are fine, so I know the quart jars from the g2g transfer had spores, so I'm wondering how long until the jars would have turned green.  This way I can colonize my jars long enough to know if they are contaminated before going through all the trouble of spawning them.  Also, I've never seen trich before now, this is the first time, and it was from the g2g transfer.  I did blast the entire lab with bleach, lysol, iso, oust even though the trays were airtight with micro-pore covered holes, and they were never opened in the lab, contents buried outside, treys filled with bleach water, cleaned, and returned to the lab. 

So, anyone know how long trich spores take to turn green on wbs?  I'm guessing 30 days since thats about the duration between the g2g transfer and when the green appeared in the 2 trays done with the g2g spawn.

Thanks again,


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InvisibleEntheogenicPeace
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Re: First trich contam, quick question on trich [Re: bbox244]
    #14728162 - 07/06/11 08:32 PM (12 years, 7 months ago)

---


Edited by EntheogenicPeace (01/09/22 07:01 PM)


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Invisiblebbox244
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Re: First trich contam, quick question on trich [Re: EntheogenicPeace]
    #14730962 - 07/07/11 12:27 PM (12 years, 7 months ago)

Correct, but I didn't actually see the trich in the jars.  I know it was there though, since the only trays that got trich were the ones which were spawned using quart jars which were inoculated with grains via g2g, not GLC which is what I used to inoculate the other jars, which were spawned to the same substrate and all of those trays are just fine.  So, I know I got the trich during the g2g transfer.  So, for next time, I'm wondering how long it takes trich to turn visibly green in a WBS jar?

The whole time from g2g to spawn, to green trick was about 30 days...  Is that about how long it would take trich to show up in spawn?  I am willing to let my quart spawn jars to colonize for 30 days to make sure they aren't infected before I go through all the trouble of making trays.  I'm just curious how trich actually "works", it's very difficult to get accurate information on time lines, etc with trich.  The only things I've found on the boards are:

-Bury everything contaminated with it.
-It turns green when it is sporulating
-Sterilize everything after an outbreak
-Pasteurization kill trich spores
-It starts off as white looking mycelium
-It won't grow on coir

And thats pretty much it, I want to know how exactly long does it take to go from spore -> turning green.  Thats the $60,000 question...  Anyone know???  Is it different for different mediums??

This way I can let my jars colonize long enough to know if they get contamed before going through all the trouble of spawning them.  I don't mind waiting an extra week or 2 if it'll save me a lot of problems. 

Or, am I looking at this all wrong?  I have a lot of book, forum, agar, wbs, cake knowledge, and hands on work, but my bulk "hands on" is severely lacking.  I've only actually done a few bulk grows.  Tons of agar, cake, and wbs work though.


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Offlinewally_world
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Re: First trich contam, quick question on trich [Re: bbox244]
    #14730996 - 07/07/11 12:34 PM (12 years, 7 months ago)

Trich will, in fact, grow on coir.

And after enough encounters with it you can catch it before it turns green as it starts with a denser, grainier, and brighter white cell mass before it sends out the green spores. This is what you want to be able to do so you can get rid of it before sporulation.


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Offlinewally_world
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Re: First trich contam, quick question on trich [Re: wally_world]
    #14731011 - 07/07/11 12:38 PM (12 years, 7 months ago)

Also, trich has a bitter smell when compared to myeclium.

And as far as your jars being infected, you would have known before you made the transfer, so I'm guessing you got contammed in your transfer process, are you confident in your sterile techniques?


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Invisiblebbox244
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Re: First trich contam, quick question on trich [Re: wally_world]
    #14731067 - 07/07/11 12:53 PM (12 years, 7 months ago)

RR says it won't grow on coir.  But, I have no idea how old that post was.  If it does grow on coir, then perhaps it wasn't the g2g, but just dumb luck that only the g2g trays got contamed, while the GLC trays are fine.  I say that because I left out the fact that the g2g trays are 1:3 and the GLC trays are 1:1. 

Either way, is there a link or a book ref, you have to showing what it looks like before it sporulates?  Can it be spotted in grain spawn jars?

Also, if you please, how long does trich take to go from spore->green?

Thanks for that, I will definitely start looking for references to early identification.

Oops, I just saw your second post pre-submission:

1) I am confident in my sterile technique, but, on that same day, I did an agar inoculation which resulted in a cobweb mold half-pint, my first cobweb experience as well, so I think on that exact day, I wasn't misting my GB with iso enough.  I traced the cobweb back to the agar plate, which had a crack in the parafilm where the cobweb originally was (I marked the crack with a sharpie) but, when I looked at the plate (tri-section) I found that the cobweb had moved onto the other sections, which btw, inoculated 2 other half pints which did NOT get cobweb.  But, the cobweb did move from the one section to the other 2 sections on the single plate after transfer.  I usually do more misting between work batches in my GB, except for that particular day.  I wasn't nearly as anal as I usually am on that day.

2) I always smell my jars, and the 1/2pint master used for the g2g smelled really strong (like mushrooms).  Also, when I spawned the quart jars I did not smell them, big mistake.  Which in retrospect, was really stupid since I do spawning in open air in a hepa filtered tight working space.  So, I could have easily done it, but now that you've mentioned it, I didn't bother, for some stupid reason.  I would've remembered if I did.  That was plain stupid.  I didn't even think about that until you said it.  Damn, the other trays could be contamed as well, and just not presented yet.  They are fully colonized, gonna add casing tonight/tomorrow, and I will definitely give them a good whiff before proceeding.


--------------------
.... Always curious, always looking for better ways, mind always wonders ....

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Edited by bbox244 (07/07/11 01:10 PM)


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Invisiblebbox244
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Re: First trich contam, quick question on trich [Re: bbox244]
    #14731122 - 07/07/11 01:07 PM (12 years, 7 months ago)

One, other thing, I also smell my GLC jars after usage to make sure they have a strong mushroom smell. 

My GLC Process:
I just take a colonized half-pint hit it against a tire to break it up, inject 60cc's and shake.  Then I draw a few CC's (per quart) and noc up 8-10 quarts, or if experimenting, I'll just use a couple CC's of water and noc up a single cake.  This is how I usually noc up my quarts, always worked in the past with no trich, by I wanted to see how fast a g2g would colonize in comparison, and I found btw that the GLC blows away the g2g.  But, the bad part of my GLC method is that I almost always lose the GLC jar after usage, sometimes, if I get enough water out, it will re-colonize and I can use it again.  That only happens like 3 out of 5 times however, if using it on cakes, which is what I usually do.  Like I said, my bulk knowledge is limited since I've been doing mostly experiments for a project I'm working on, this was simply to make a batch for an awesome tincture method I want to try out (gonna try and extract both primary actives cin&cybin).


--------------------
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InvisibleEntheogenicPeace
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Re: First trich contam, quick question on trich [Re: bbox244]
    #14731226 - 07/07/11 01:31 PM (12 years, 7 months ago)

---


Edited by EntheogenicPeace (01/09/22 07:01 PM)


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Offlinewally_world
Mr. Cook

Registered: 02/01/09
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Re: First trich contam, quick question on trich [Re: bbox244]
    #14731482 - 07/07/11 02:26 PM (12 years, 7 months ago)

Quote:

bbox244 said:
RR says it won't grow on coir. 

Either way, is there a link or a book ref, you have to showing what it looks like before it sporulates?  Can it be spotted in grain spawn jars?

Also, if you please, how long does trich take to go from spore->green?





I have had trichoderma bloom on coir before, but come to think of it, I did have other ingredients mixed in with the coir. However, Coir is used as growing media for plants and there are plant-beneficial tichoderma species you can buy to "feed" your plants which causes a symbiotic relationship between plant and the trich bacteria. Therefore, I can't see a reason trich would NOT grow on coir. Hopefully RR would like to chime in.

Here's a link to a pic of trich on agar plates, one sporulated and one blooming.
http://www.biocontrol.entomology.cornell.edu/pathogens/trichoderma.html

As far as a timeline goes, it's really hard to say as trich is somewhat similar to myc in that it will sporulate when it feels threatened. This could mean it has colonized all the available nutrients, ran into a competitor organism, or even experienced threatening environmental factors. So as far as a timeline goes, there really isn't one, which is why you must learn to spot the trich "mycelium" before it turns green. And trust m, it gets easy to do.

Quote:

bbox244 said:
1) I am confident in my sterile technique, but, on that same day, I did an agar inoculation which resulted in a cobweb mold half-pint, my first cobweb experience as well, so I think on that exact day, I wasn't misting my GB with iso enough.





So you are doing agar work in a GB? This is a problem in itself. GB's do not provide the stringent filtration needed to properly perform the sterile techniques needed for agar. You really need a flowhood. However, if you're still intent on doing agar work in a GB, get some media antibiotic and/or look into some antibiotic agar recipes.


Quote:

bbox244 said:
2) I always smell my jars, and the 1/2pint master used for the g2g smelled really strong (like mushrooms)... They are fully colonized, gonna add casing tonight/tomorrow, and I will definitely give them a good whiff before proceeding.




Smelling is NOT the most efficient way to detect trich as you really don't want your mouth/nose/face anywhere NEAR an exposed spawn/substrate/agar plate, etc.. Humans have very high bacterial counts in their mouths and you don't want that close to items you want to remain sterile. This is why you must learn what trich looks like, not to be a broken record or anything.

However, smelling can be useful when trying to detect trich in a colonizing casing, or even better a mycobag (just squeeze the bag and smell the air coming from the filter-patch), b/c you can keep your nose/mouth/breath AWAY from the goodies. But as for smelling jars/plates/LC's before inoculation, BIG NO-NO! :smile:.


Quote:

bbox244 said:

My GLC Process:
But, the bad part of my GLC method is that I almost always lose the GLC jar after usage, sometimes, if I get enough water out, it will re-colonize and I can use it again.




Yeah, you really don't want re-use them, anyhow. What you can do, though, is inject distilled water multiple times PER SESSION. However, injecting a bunch of water into a jar of grain and letting it sit for a few days is never a good thing, even if you manage to get MOST of the water out of there.


Lastly, yes you can spot trich on grain. You can spot it wherever it's growing if you know what to look for.

Here are a few signs to look for (sorry there's no pics):
1.) If you see a jar/casing/bag/plate that isn't doing as good as the rest in the batch, start looking.
2.) Stalled spawn/substrate
3.) Bright white (brighter than myc) cell mass that has a grainy texture to it (like in the pic linked to above)
4.) The bitter smell

Hope this helps!


--------------------
^False and fictitious.


Edited by wally_world (07/07/11 02:27 PM)


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Invisiblebbox244
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Re: First trich contam, quick question on trich [Re: wally_world]
    #14751864 - 07/11/11 03:01 PM (12 years, 7 months ago)

Thank you very much for the info.  There are some really solid answers in there.  When I take a whiff, I just move my surgical mask down enough to smell, I am very very careful.  I have a great sense of smell and I can simply waft my iso soaked glove over the top of a jar to smell for the sour smell, but I agree, this isn't the way I want to detect trich.  It's great information to know that the times it takes for trich to sporulate varies.  Since the time isn't fixed, I will spend some time researching identification of it before it sporulates, and puts my lab at risk.  I do use nothing but sealed plates, jars, trays, but nonetheless, I don't want any sporulating trich in my lab, period. 

I did see in the pictures the "grainy" type of growth trich displays.  I would imagine it's density is probably greater than mycelium's as well being it's parasitic.  So perhaps this is another way to tell in grains. 

As far as using a GB, those days are now over since I received my HEPA filters.  My HEPA laminar flow hood will be up and running in a few days, so I won't have anymore GB disasters.  I've done hundreds of plates in my GB, and I have had some contaminates in the past from particles floating around inside my GB, so all of that will be gone now, thank goodness.

Nonetheless, since I've now seen it for my first time, I'm very paranoid about it.  Sub pasteurization could have been a contributing factor as well, but not for next time.  I have taken my pasteurization bucket and covered it with radiant insulation.  I did a little experiment to see how hot water would stay hot in the bucket with insulation and the difference was tremendous.  So, I'm going to use the insulated bucket next time to ensure everything in there gets fully pasteurized completely.


--------------------
.... Always curious, always looking for better ways, mind always wonders ....

TRUE HEPA Filtered Air Pumps TEK
Everything you need to know about tubs TEK
Fast Tub Dehydrator TEK


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Offlinewally_world
Mr. Cook

Registered: 02/01/09
Posts: 171
Last seen: 5 years, 3 months
Re: First trich contam, quick question on trich [Re: bbox244]
    #14752984 - 07/11/11 06:48 PM (12 years, 7 months ago)

Yeah you made the best decision of your mycological life in obtaining a flowhood. I messed around with still-air inoc's and spraying oodles of oust (and still getting contams) for 2 years. Then I got a flowhood and kicked myself in the ass for not getting one sooner. Their initial cost is a bit high but it is more than compensated for within a couple of uses, IMO, when you consider the cost of oust and wasted time/money on contaminated media.


Good idea on insulating the bucket. I'd say that will do you just fine but if you ever get to being a total past.-temp. control freak like me, then try a large turkey fryer. I found 2 35-quart turkey fryers on clearance at lowe's for $25 a piece and they are very convenient, although may be a bit overboard. But I do like the fact that if the temp drops all I have to do is turn on the flame vs. keeping an extra pan of boiling water ready on oven. They also work great for the simple task of hydrating coir. However, I wouldn't advise buying one at full price as they can be expensive and will have a rough ride when purchased for mycological intentions.


Lastly, although I'm sure you're aware, never opened contammed bags/trays/jars/plates inside any part of your house. Especially with trich, once you see the green, there millions of spores jumping to ruin your day.


--------------------
^False and fictitious.


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