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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
trich in fully colonized wbs jars?
    #18800482 - 09/04/13 06:43 PM (10 years, 5 months ago)

I've been cultivating for six or seven months now, and recently tried by first wbs g2g transfer with P. cubensis Tasmanian.  I used a glove box, wiped down everything with alcohol, gloves, long sleeves, surgical mask.  At first the results appeared too good to be true... a bit of green (trich?) in only one of the seven the receiving quart jars.  After 8 days, all of the jars are 99-100% colonized, including the one that I thought was contaminated with trich.  I thought it was out-competed or I was just being paranoid and it was never there.  But now that the jars are at 99-100% colonized, it appears that some of them may have the faintest hint of green in the mycelium.  Could I have achieved 100% colonization even if there is trich, or some other mold contaminant, in my jars?  Even if they are contaminated, can I go ahead and use them for spawning a bulk substrate (perhaps with some crushed rhododendron leaves)?  They look so pretty and healthy, although some of them definitely look like they may have trich.


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Offlineincubescence
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Registered: 02/05/13
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Last seen: 9 years, 3 months
Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18800523 - 09/04/13 06:54 PM (10 years, 5 months ago)

Best advice is to take a pic.  It sounds like the first one is probably but the others sound more like bruising.  Other than that I'd say let them look at it  :smile:

And I'm sticking this in my signature.

How it should & shouldn't look - NEW CULTIVATORS GUIDE


Edited by incubescence (09/04/13 06:58 PM)


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: incubescence]
    #18800951 - 09/04/13 08:49 PM (10 years, 5 months ago)

I've attached pics, although I don't think they'll help.  In jar one, there are a few kernels in the foreground that look like they might have a gritty green substance.  That's the jar that I initially thought was contaminated, but it continued to colonize nicely.  In jar 2, just below the '800' mark, under the surface of the mycelium, is some faint blue/green discoloration, but you can't see it in a pic.  Again, the jar looks well colonized overall (there is a huge chunk of colonized seed at the top of jar 2... it fell in during the g2g transfer). 

I didn't think pics would help much, except to show that they are fully colonized, regardless of any potential contaminants...


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18801493 - 09/04/13 10:59 PM (10 years, 5 months ago)

Perhaps the blue spots are bruising...  It was most noticeable just after shaking the mother jar, and then noticeable today, when I shook one of the colonized receiving jars to spawn a new generation.  I read that shaking colonized grain can cause bruising?

In any event, my main question is, would a jar colonize 100% and look pretty, without any alarming defects, and still be harboring troublesome molds (eg, trichoderma, aspergillus) that will manifest and take over uncolonized grain when they're used in a G2G transfer?

Thanks for any help.


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OfflineMAIA
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Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18801533 - 09/04/13 11:10 PM (10 years, 5 months ago)

Yes, shaking the jars can cause bruising. Give it a couple of days. Bruising doesn't spread and maintains a regular coloration. "Good" colors are blue or grayish blue. There could be a greenish hue on that blue if heavily bruised. Be patient, wait and see ...


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Spiritual being, living a human experience ... The Shroomery Mandala



Use, do not abuse; neither abstinence nor excess ever renders man happy.
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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: MAIA]
    #18803128 - 09/05/13 12:12 PM (10 years, 5 months ago)

I think jar 1 definitely has a contaminant, but I think the others are bruising.  In jar 1, two of the seeds clearly have a green sandy powder on them.  The funny thing is that it's been there for over a week now, yet the jar continued to reach 100% colonization, minus those two kernels.  Why isn't the contaminant mold spreading?  The other jars are probably bruising, but jar 1 definitely looks like it has mold spores.  Shouldn't that mold have taken over the entire jar by now?  And should I go ahead and use it for spawning a bulk substrate?

Thanks again everyone for your help.  I've had so many wbs jar contams lately, I think it's from pushing the spore syringe through my polyfill, then the polyfill gets wet, thus allowing contaminant introduction.  But it seems like I'm having better luck with G2G, because I open the lids (in a glove box, with all the precautions I can think of) and then close them, without affecting the integrity of my filters.  I have a filter setup which I'm beginning to think is no good.  I use a piece of tyvek, topped with a metal lid with a 1/4" hole stuffed with polyfill, and I stick a syringe through that hole.  Often, the polyfill is pushed through the tyvek, then gets wet, and prob introduces contams.  I will change this filter arrangement soon.  My contam issues have decreased a lot by: not allowing kernels to explode, moving the inoculation process out of the kitchen, and wearing long sleeves and a surgical mask (in addition to isopropanol, gloves, and glovebox.)

Thanks all.


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18818558 - 09/09/13 11:20 AM (10 years, 5 months ago)

I never figured out what the contaminant was in jar 1 (grainy, powdery green substance covering two kernels at the top of the jar), but the mushroom mycelium has since grown over it!  I can't see any signs of contaminants now.  I guess whatever it was, the mycelium won out.  I think I will try to use it in a bulk substrate and see what happens.


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OfflineKizzle
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Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18818816 - 09/09/13 12:51 PM (10 years, 5 months ago)

Quote:

Why isn't the contaminant mold spreading?



Probably because you're looking for green. The mold itself is white, only the spores it produces, where and when it decides to produce them, are green. That's not say mushroom mycelium can't out-compete any molds but it's probably wishful thinking. Since you know there's mold it I would suggest fruiting it outdoors both for a better a chance of it successfully fruiting despite the presence of mold and to prevent contaminating your grow areas.


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InvisibleDreaming Nomad
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Registered: 01/15/13
Posts: 908
Re: trich in fully colonized wbs jars? [Re: Kizzle]
    #18821417 - 09/09/13 11:40 PM (10 years, 5 months ago)

Look for a mass of white mycelium spheres that resembles a cluster of hyphal knots. This is what trich myc looks like. It's easy to overlook it or mistake it for cubensis, but once you see it you'll never forget. The green stuff will appear over that soon afterwards. By then its probably too late... Trich is very aggressive, at least the trich I got was!

But then again there are many green molds/fungi out there and green doesn't necessarily mean trich.


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They say curiosity killed the cat...
Fortunately, I am not a cat.


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: Kizzle]
    #18821461 - 09/10/13 12:01 AM (10 years, 5 months ago)

Well it was definitely there at one point, but I have a feeling it might have been some kind of Aspergillus rather than trich. I saw a pic of Aspergillus spores that looked like little grains of green sand.  Actually I think I've misidentified Aspergillus as trichoderma several times.  Usually when I find this contaminant in a jar, it spreads all throughout, but this time it didn't.  I would really like to correctly identify this contaminant; I may take a sample up to the local university and have a look under a microscope.  It's been years since I've taken microbiology, but my old micro professor would be glad to help identify it.

And you're probably right, even if the mushroom mycelium was able to colonize those two contaminated kernels, I probably shouldn't take a chance by using that jar indoors.  I'll use it outside.

Those were some great contam pics; thanks.


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18821477 - 09/10/13 12:07 AM (10 years, 5 months ago)

Hey nomad,
the jar was inoc'd 2 weeks ago and they look great now.  I'm starting to think that it wasn't trich after all, since it hasn't taken over the jar. But like you said, it could be one of many molds, and apparently it's one that isn't as aggressive as P cubensis.  I'll post what happens.


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InvisibleDreaming Nomad
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Registered: 01/15/13
Posts: 908
Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18821489 - 09/10/13 12:11 AM (10 years, 5 months ago)

Yeah let us know what happens!


--------------------
They say curiosity killed the cat...
Fortunately, I am not a cat.


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OfflineKizzle
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Registered: 08/30/11
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Re: trich in fully colonized wbs jars? [Re: Dreaming Nomad]
    #18824497 - 09/10/13 05:53 PM (10 years, 5 months ago)

Quote:

Look for a mass of white mycelium spheres that resembles a cluster of hyphal knots.



Actually that sounds more like Aspergillus spp. Aspergillus looks a lot like mushroom mycelium but produces little pins that turn green, yellow or black. Trichoderma generally produces a thick bubbly shaped mycelium that turn green after spore production.


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: Kizzle]
    #18830261 - 09/11/13 10:00 PM (10 years, 5 months ago)

Yeah, I think you're right Kizzle, I think it's an Aspergillus species.

I'm really tempted to put it in a bulk substrate indoors (far from the others) just to see what happens, but I know it would be best just to use it outdoors.  In any event, it's beautifully colonized...


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? [Re: dukeh98]
    #18859624 - 09/18/13 06:23 PM (10 years, 5 months ago)

I used the jars in a bulk substrate and the mycelium has been colonizing just fine...


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Offlinedukeh98
mycologist wannabe

Registered: 07/23/13
Posts: 19
Last seen: 10 years, 3 months
Re: trich in fully colonized wbs jars? UPDATE [Re: dukeh98]
    #18997125 - 10/18/13 07:42 PM (10 years, 4 months ago)

These jars produced three large flushes in a bulk substrate before eventually being overtaken by contaminants.  I have one jar that I never used for spawn... fruits are growing inside the jar.  After over a month later, the mycelium is still quite healthy and there are no more signs of the contaminant.  In the future I will use jars that seem to 'out-compete' contaminants only if I don't have healthier alternative jars.  But they worked just fine!


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Invisibleenolagay
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Registered: 01/16/14
Posts: 3
Re: trich in fully colonized wbs jars? UPDATE [Re: dukeh98]
    #19448473 - 01/20/14 04:27 PM (10 years, 1 month ago)

The jars are fine if you go with Ball usually... Also I find silicone gel a lot more productive than polyfil for a inoculation port.  It seals extraordinarily well and can go through multiple pressure cooks before being beat.  Good news with your flushes though, and keep it up


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