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Odd Clusterfukc
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Fuzzy agar and 3 different looking LC - first timer
#27312776 - 05/17/21 04:30 PM (3 years, 7 months ago) |
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Hello people, I'm a newbie here, came to show you some of my first attempts with both agar and LC and maybe get some directions too 
Sooooo, made some agar and dropped some spores from a print just about a week ago. All dishes show now similar growth... a thin white fuzz. I know being a multispore I'm expected to see fluff more than anything else, just wondering if that is the right fuzz or not lol but you can be the judge of that.
 Should I just transfer and see?
I've also started some LC (3% honey). I've made 3 jars and inoculated each one of them in a different way.
 A) injected with a MS syringe made by me a couple days earlier. Checked the jar within 3 hours from injection and already I couldn't see any spores but tiny white specs. Now I'm at day 7 and LC got cloudier with "dandruff" as in the pic. It sits on the bottom until swirled. I exclude it's sediments because the other jars have none and they were all made at the same time.
 B) scraped in some spores from the same print I made the syringe with. Some of the spores seem now to have germinated. It's got tiny specs and fluff floating about. Seems fine to me
 C) last one was inoculated with tissue from a stem. It's now started to grow on the edges. This seems ok too but I might be wrong. Picture isn't great sorry, I tried my best for 15 minutes. Man, I never knew getting a decent picture of this stuff would be so challenging lol
So that was it. Any thoughts? I'm mostly dubious about the agar and the jar A... but I guess experience ey
Edited by Odd Clusterfukc (05/17/21 04:34 PM)
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Tormato  
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: Odd Clusterfukc]
#27312943 - 05/17/21 07:22 PM (3 years, 7 months ago) |
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A, B, and C will all have a high probability of contams. Shooting spores, dropping mushroom tissue or scrapping spores directly to LC mix is a horrible crap shoot.
As far as your plates go they look ok for transfers.
The best route for making LC is to use a clean culture from agar.
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Odd Clusterfukc
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: Tormato]
#27312999 - 05/17/21 08:21 PM (3 years, 7 months ago) |
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Thank you for the feedback  About the LC yes, I'm aware of that. I guess I'll only find out with a test jar or trying it out on agar. Even though in the end, if I used the same spores to start both agar and LC, I reckon the chances of contam are exactly the same. The only difference is that I might not be able to spot it in LC
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HerbalSteve
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: Odd Clusterfukc]
#27313078 - 05/17/21 10:40 PM (3 years, 7 months ago) |
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No not remotely. Liquids allow for maximum dispersion throughout. You go to agar, 2 - 3 transfers later you can start to think about making LC/LI. Honestly you'll get a much better result and response than just shooting your load into some bacterial broth.
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Odd Clusterfukc
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: HerbalSteve]
#27313227 - 05/18/21 03:35 AM (3 years, 7 months ago) |
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Quote:
HerbalSteve said: No not remotely. Liquids allow for maximum dispersion throughout. You go to agar, 2 - 3 transfers later you can start to think about making LC/LI. Honestly you'll get a much better result and response than just shooting your load into some bacterial broth.
What do you mean "not remotely"? I understand the dispersion problem. Stuff would start floating about and potentially germinate freely anywhere in the solution. But if I scraped the same contaminated spores in agar and LC under the same *as sterile as possible conditions*, wouldn't they both end up contaminated?
Edited by Odd Clusterfukc (05/18/21 07:41 AM)
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Mateja



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Re: Fuzzy agar and 3 different looking LC - first timer [Re: Odd Clusterfukc]
#27313253 - 05/18/21 04:50 AM (3 years, 7 months ago) |
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Quote:
Odd Clusterfukc said: if I scraped the same contaminated spores in agar and LC under the same *as sterile as possible conditions*, wouldn't they both end up contaminated?
Bacteria grows poorly on agar you might not even notice it, and in liquid media bacteria can not hide it will either make the broth turbid overnight or it will not even present itself at all which means it's not present.
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Odd Clusterfukc
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: Mateja]
#27313286 - 05/18/21 05:42 AM (3 years, 7 months ago) |
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Quote:
Mateja said: Bacteria grows poorly on agar you might not even notice it
Ah that's cool I didn't know. I was under the impression that contams in LC can hide very well though, and that's why it's recommended to go to agar first
Edited by Odd Clusterfukc (05/18/21 07:23 AM)
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sandman420
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: Mateja]
#27313287 - 05/18/21 05:42 AM (3 years, 7 months ago) |
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Yes you are getting great info here. Contams will 100% germinate that get in LC and permeate it. A few little contam spores may or may not germinate on agar before they are destroyed by mycelium and those that do can be seen and avoided.
Tissue, spores, grains, all that stuff is really risky to put to LC. Sure it can be done, but even the best grower would be looking at a percentage of failure with those methods.
Edited by sandman420 (05/18/21 05:45 AM)
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FreshStart
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: sandman420]
#27313306 - 05/18/21 06:06 AM (3 years, 7 months ago) |
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I'm surprised no one has said with agar you can ALWAYS transfer clean looking mycelium to another agar plate. It's practically the steps leading up to inoculating grain or LC. I transfer early and often until I'm comfortable with what I have on agar then introduce it to other growth medium (about 3 transfers off the original plate). Also, mycelium can be rhizomorphic or tomentose, stringy or fuzzy. Good luck!
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Odd Clusterfukc
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: sandman420]
#27313393 - 05/18/21 07:27 AM (3 years, 7 months ago) |
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Quote:
sandman420 said: Yes you are getting great info here. Contams will 100% germinate that get in LC and permeate it. A few little contam spores may or may not germinate on agar before they are destroyed by mycelium and those that do can be seen and avoided.
Tissue, spores, grains, all that stuff is really risky to put to LC. Sure it can be done, but even the best grower would be looking at a percentage of failure with those methods.
Thank you for clearing that up. How is it that contams in LC can hide so well though?
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Odd Clusterfukc
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: FreshStart]
#27313409 - 05/18/21 07:39 AM (3 years, 7 months ago) |
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Quote:
FreshStart said: I'm surprised no one has said with agar you can ALWAYS transfer clean looking mycelium to another agar plate. It's practically the steps leading up to inoculating grain or LC. I transfer early and often until I'm comfortable with what I have on agar then introduce it to other growth medium (about 3 transfers off the original plate). Also, mycelium can be rhizomorphic or tomentose, stringy or fuzzy. Good luck!
That is true, that's why I can't wait1 to start the transfers. That was my first LC, that I was planning to put to agar anyway. If they're contaminated at least I might find out what to look for when looking for contams in LC (if I'm lucky lol).
Why do you transfer early? To avoid different growths touching each other? I've just read in a different thread that letting plates overgrow can give nice hyphae knots, which when transferred will grow into nice rhyzomorphic myc. They were talking about clones though...
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FreshStart
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: Odd Clusterfukc]
#27314622 - 05/19/21 05:45 AM (3 years, 7 months ago) |
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Try not to look too far ahead, you may get some info mixed up. Your LC will almost 100% be contaminated. If you put some contaminated LC onto agar it will be bacterial and grow mold because all the bad stuff already germinated in your LC. You start with agar because it's decently resistant and usually the first thing to grow will be myc, that's why I transfer early. I want to take the healthy myc edge to a new plate before the first plate grows anything else, that's not saying that it'll be a clean transfer, bacteria and mold can be tricky and in rare occasions(from my experience) and survive many transfers only to germinate later on in a jar. To see what contaminated LC looks like just use the search bar, if you click "search our forums" you get more advanced search options. As far as getting knots in a plate, it happens. If you have a clean plate and leave it for about a month you'll get pins growing but that doesn't guarantee the characteristics of your plate will be what you're looking for, too many things to take into account. Hope this helps!
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Odd Clusterfukc
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Re: Fuzzy agar and 3 different looking LC - first timer [Re: FreshStart]
#27314635 - 05/19/21 06:08 AM (3 years, 7 months ago) |
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Quote:
FreshStart said: Try not to look too far ahead, you may get some info mixed up. Your LC will almost 100% be contaminated. If you put some contaminated LC onto agar it will be bacterial and grow mold because all the bad stuff already germinated in your LC. You start with agar because it's decently resistant and usually the first thing to grow will be myc, that's why I transfer early. I want to take the healthy myc edge to a new plate before the first plate grows anything else, that's not saying that it'll be a clean transfer, bacteria and mold can be tricky and in rare occasions(from my experience) and survive many transfers only to germinate later on in a jar. To see what contaminated LC looks like just use the search bar, if you click "search our forums" you get more advanced search options. As far as getting knots in a plate, it happens. If you have a clean plate and leave it for about a month you'll get pins growing but that doesn't guarantee the characteristics of your plate will be what you're looking for, too many things to take into account. Hope this helps!
Thank you for your reply that makes sense. It's actually the first time I hear that ungerminated contams might hide in agar too, especially after many transfers. That's helpful though.
I did look for contaminated LC pics, but a part from the obvious ones I keep reading they will look exactly like healthy ones
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