|
Biscuits



Registered: 06/29/17
Posts: 162
Loc: In a society
Last seen: 2 years, 7 months
|
Spotting Hidden Trich on Agar?
#26829918 - 07/17/20 11:11 PM (3 years, 6 months ago) |
|
|
Hi,
My question will be about how to spot hidden trich on an agar plate, from suspected but not confirmed contaminated spawn. And if not in the spawn, is it possible that coir carried the trich.
On the first day of the month I spawned 6 quarts of rye (all from one agar plate) to 5 (one jar was bacterial) shoeboxes with coir using the cooler tek (coir + boiling water in a cooler left overnight), done slightly under field capacity with a soaked top layer of coir (using cooled boiled water).
Today I have buried 4 young tubs before their time, as they all spontaneously sprouted up trich overnight, and no doubt the 5th will follow soon (I think I can spot some starting). I have 2 other shoeboxes from different spawn / coir (one with casing, one without), that both do not seem to have any trich in them yet.
I also had a big monotub running that was starting to fruit (late, 1 month old, but still fruiting), that actually showed trich a day before the shoeboxes...
Which came first I do not know, but it is odd that every shoebox from that one plate / same batch of coir threw out trich and the others did not. I should also mention that the bacterial jar I threw out and covered with leftover coir also got trich outside.
Now everyone seems to say "trich is pretty much always bad spawn", so I'm starting here. I actually took a grain from one of the jars that eventually got the trich -- can anyone spot any trich or any red flags?
2 days since agar (1% agar):

4 days (the marks are just from soap water in the SAB):

6 days:
 Note that it's very fluffy, but I assume that's because it was from a grain... and that whole run was very tomentose...

I then transferred it to some more grain to see what would happen... Can anyone spot any hidden trich? Is it even possible to spot it like this?? I don't want to have to bury any more tub-children before they get to grow up.
Edited by Biscuits (07/18/20 12:00 AM)
|
w00tmycelium
Stranger


Registered: 11/28/19
Posts: 159
Last seen: 3 years, 4 months
|
Re: Spotting Hidden Trich on Agar? [Re: Biscuits]
#26829929 - 07/17/20 11:22 PM (3 years, 6 months ago) |
|
|
Based on those plate pictures, they seem perfectly fine. Some decent rhizo patterns underneath, normal tomentose on top, also very uniform going outward... I would just wait, because the plates seem fine currently to me. If they are contaminated somewhere in some segment, sure, slice them out and transfer but no big deal.
|
Bsdgaou

Registered: 02/20/19
Posts: 340
|
Re: Spotting Hidden Trich on Agar? [Re: w00tmycelium]
#26829939 - 07/17/20 11:33 PM (3 years, 6 months ago) |
|
|
Yea, look nice.
|
southbounpachyderm
Stranger
Registered: 07/20/18
Posts: 282
Last seen: 3 years, 5 months
|
Re: Spotting Hidden Trich on Agar? [Re: w00tmycelium]
#26829941 - 07/17/20 11:33 PM (3 years, 6 months ago) |
|
|
Are these the plates you used? If you have any pictures of the plates you used that would be super helpful. Your spawn might have been bacterial which can weaken the myc to the point that trich can get a foothold. I normally see my plates, even if its multispore, about half way or more thru the plate by day 6. Normally takes 10-12 days from transfer to fully colonized plate. When I see fluffy slow growth like this I immediately assume bacteria are present. Might be your issue here.
|
w00tmycelium
Stranger


Registered: 11/28/19
Posts: 159
Last seen: 3 years, 4 months
|
|
Quote:
southbounpachyderm said: Are these the plates you used? If you have any pictures of the plates you used that would be super helpful. Your spawn might have been bacterial which can weaken the myc to the point that trich can get a foothold. I normally see my plates, even if its multispore, about half way or more thru the plate by day 6. Normally takes 10-12 days from transfer to fully colonized plate. When I see fluffy slow growth like this I immediately assume bacteria are present. Might be your issue here.
Going to disagree here, because I see so much good fluffy growth on top that it's insane compared to the rhizo underneath. I have a theory that the fluffy parts are just the colony trying to reach more nutrients/substrates. I've seen similar behavior directly from spore to grain, as well as agar to grain. Just depends on the colony/genetics from what I can tell.
|
Biscuits



Registered: 06/29/17
Posts: 162
Loc: In a society
Last seen: 2 years, 7 months
|
|
I think it would have been one of these, although I can't remember which (all three of these are transfers from the same plate). Unfortunately I did not take any back shots.

The "wave" that's in there I attributed to a few hours of cold that I subjected them to while doing some renovations -- do you think it could be something else?
Edit: These plates are 7 days old using 4% agar
Edited by Biscuits (07/17/20 11:59 PM)
|
Biscuits



Registered: 06/29/17
Posts: 162
Loc: In a society
Last seen: 2 years, 7 months
|
Re: Spotting Hidden Trich on Agar? [Re: Biscuits]
#26829975 - 07/17/20 11:56 PM (3 years, 6 months ago) |
|
|
Also i should mention that the grain plate I used a mere 1% agar, compared to those three where I used 4%.
|
w00tmycelium
Stranger


Registered: 11/28/19
Posts: 159
Last seen: 3 years, 4 months
|
Re: Spotting Hidden Trich on Agar? [Re: Biscuits]
#26829982 - 07/18/20 12:01 AM (3 years, 6 months ago) |
|
|
Quote:
Biscuits said: I think it would have been one of these, although I can't remember which (all three of these are transfers from the same plate). Unfortunately I did not take any back shots.
The "wave" that's in there I attributed to a few hours of cold that I subjected them to while doing some renovations -- do you think it could be something else?
Edit: These plates are 7 days old using 4% agar
Those look great to me. Blurry pictures but still great.. uniform growth outward. Also great work on the transfer, good uniform square.
|
southbounpachyderm
Stranger
Registered: 07/20/18
Posts: 282
Last seen: 3 years, 5 months
|
Re: Spotting Hidden Trich on Agar? [Re: w00tmycelium]
#26829984 - 07/18/20 12:02 AM (3 years, 6 months ago) |
|
|
Thats absolutely not my experience with agar. Normally slow fluffy raised growth is bacterial. Disagree all you want this is simply my experience throughout thousands of plates and hundreds of tubs at this point.
|
southbounpachyderm
Stranger
Registered: 07/20/18
Posts: 282
Last seen: 3 years, 5 months
|
Re: Spotting Hidden Trich on Agar? [Re: Biscuits]
#26829992 - 07/18/20 12:06 AM (3 years, 6 months ago) |
|
|
Perfect. Those plates are 100% bacterial. Do you see that ring around your myc that looks like a little halo and has a different appearance. That's an indicator of bacterial contamination.
Also, less nutrients means the mycelium grows more quickly searching for more food. It slows down when more nutrients are present which is another good indicator your grain plate is in fact bacterial as well.
Edit: 4% nutes at 500ml of water is 20g of lme. thats wayyyy too much. typical recipe is 10g lme per 500ml of water. I personally prefer 7g's lme to 500 ml water.
Edited by southbounpachyderm (07/18/20 12:11 AM)
|
Biscuits



Registered: 06/29/17
Posts: 162
Loc: In a society
Last seen: 2 years, 7 months
|
|
Ah this is excellent news, I now probably know where the problem is which is 90% of the battle I think. It seems like it was bacterial, that weakened it and the trich in the air jumped on the opportunity... Right?
I was thinking that the grain one might be slow and tomentose because it has nutrients from the grain itself? Or maybe that doesn't matter.
So how do I isolate away from the bacteria if it is in a halo around the myc?
|
southbounpachyderm
Stranger
Registered: 07/20/18
Posts: 282
Last seen: 3 years, 5 months
|
Re: Spotting Hidden Trich on Agar? [Re: Biscuits]
#26830010 - 07/18/20 12:24 AM (3 years, 6 months ago) |
|
|
Quote:
Biscuits said: Ah this is excellent news, I now probably know where the problem is which is 90% of the battle I think. It seems like it was bacterial, that weakened it and the trich in the air jumped on the opportunity... Right?
I was thinking that the grain one might be slow and tomentose because it has nutrients from the grain itself? Or maybe that doesn't matter.
So how do I isolate away from the bacteria if it is in a halo around the myc?
That is my best guess. Trich is a pretty quick mold to take over and sporulate if it was in your spawn or agar at the start you would have seen it happen within 3-4 days.
Personally I have very rarely had success getting rid of this type of bacterial infection. Josex's poke method is a last ditch effort if I want to save the culture. I've heard people have success with using water agar because the myc grows faster than the bacteria at that point.
as far as slow and tomentose from the grain having nutes. That is a possibility I've never dropped a grain to an agar plate so I have never watched what it does in real time. Maybe I should do that tomorrow after I spawn my tubs...
|
Biscuits



Registered: 06/29/17
Posts: 162
Loc: In a society
Last seen: 2 years, 7 months
|
|
Ah I had never heard of this Poke method before, but I have everything I need to try it so here goes! Keen to learn a new method. Thanks
|
southbounpachyderm
Stranger
Registered: 07/20/18
Posts: 282
Last seen: 3 years, 5 months
|
Re: Spotting Hidden Trich on Agar? [Re: Biscuits]
#26830026 - 07/18/20 12:37 AM (3 years, 6 months ago) |
|
|
It's definitely saved a few of my cultures before and is the only way I will do liquid cultures. Very useful method. Also, no thanks needed. I got a lot of help from the community when I started out. I am eager to share my knowledge now that I feel good enough at growing and the intricacies of everything involved in it. I can finally accurately give advice!
|
w00tmycelium
Stranger


Registered: 11/28/19
Posts: 159
Last seen: 3 years, 4 months
|
|
Quote:
southbounpachyderm said: It's definitely saved a few of my cultures before and is the only way I will do liquid cultures. Very useful method. Also, no thanks needed. I got a lot of help from the community when I started out. I am eager to share my knowledge now that I feel good enough at growing and the intricacies of everything involved in it. I can finally accurately give advice!
I will have to defer to your experience, but I am now tempted to show a ton of extremely healthy plates with fluffy growth up top. Just wish it was easier to post pics lol.
https://i.imgur.com/MD3Li.jpg
But to post this again, these came out of fluffy mycelium, still even looks fluffy on the coir by the end stages on some sections.
|
southbounpachyderm
Stranger
Registered: 07/20/18
Posts: 282
Last seen: 3 years, 5 months
|
Re: Spotting Hidden Trich on Agar? [Re: w00tmycelium]
#26830050 - 07/18/20 01:03 AM (3 years, 6 months ago) |
|
|
Tomentose doesn't always mean its bacterial. When it looks like his plates did tho in my experience it does mean bacterial presence. That's just an observation I've made.
I've been thinking of grabbing a microscope with a usb link to the computer for capturing pictures. That way I can document the different types of growth I find common with bacterial issues and the fact that they are bacterial. There are a few signs that are extremely common and good indicators.
|
w00tmycelium
Stranger


Registered: 11/28/19
Posts: 159
Last seen: 3 years, 4 months
|
|
Quote:
southbounpachyderm said: Tomentose doesn't always mean its bacterial. When it looks like his plates did tho in my experience it does mean bacterial presence. That's just an observation I've made.
I've been thinking of grabbing a microscope with a usb link to the computer for capturing pictures. That way I can document the different types of growth I find common with bacterial issues and the fact that they are bacterial. There are a few signs that are extremely common and good indicators.
That's exactly why I got the microscope, and it's easy to get pictures out of it. I just hate uploading them. But I'd recommend getting some slides, covers and dyes because it seems to help identifying the hyphae. But I've also noticed that there's some hyphae that looks healthy but is not the good kind.
|
|