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OfflineBramscoChill
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liquid culture contamination
    #26492718 - 02/19/20 01:08 AM (3 years, 11 months ago)

I created 5 jars of liquid culture and used them with the BRF TEK. It seems to go allright. I used an syringe filter of .22um and RTV as injection port.

The jars with LC are about 3 weeks old. 1 jar I injected some jars with a day ago (I use isoprop alcohol and mouth mask, cooking the needles+syringe for half an hour, heating the needles), is showing small gray spots. Also grey spots attached to the glass an the side. It got there in like 1 day.

I added some waterperoxide in case it is an contermination.

Does anyone know if it's conterminated?

pics:


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Invisiblewildernessjunkie
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Re: liquid culture contamination [Re: BramscoChill]
    #26492724 - 02/19/20 01:12 AM (3 years, 11 months ago)

Usually you need to do test jars/plates to tell if an LC is bad. Except for those, no need to test them. They are obviously bad.

Its never a good idea to inject spores to LC.


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OfflineBramscoChill
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Re: liquid culture contamination [Re: wildernessjunkie]
    #26492730 - 02/19/20 01:22 AM (3 years, 11 months ago)

Is it because of the grey spots?
How to tell if an LC is gone bad?


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Invisiblewildernessjunkie
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Re: liquid culture contamination [Re: BramscoChill]
    #26492758 - 02/19/20 02:09 AM (3 years, 11 months ago)

Most of the time, you cannot tell visually if an LC is bad. You have to test it on agar, or in some test jars first.

But that dark color ring around the top of your liquid is clearly some kind of mold.


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Invisiblec10h12n2o
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Re: liquid culture contamination [Re: wildernessjunkie] * 1
    #26492774 - 02/19/20 02:51 AM (3 years, 11 months ago)

What did you inject it with? Spores?

Spores are not sterile so its always a bad idea to make lc with spores

Also what was the point of the peroxide?


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Offlinemr. whothehell
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Re: liquid culture contamination [Re: c10h12n2o]
    #26492803 - 02/19/20 03:53 AM (3 years, 11 months ago)

almost any time you inject spore directly on LC you'll end up with contam, if we cannot tell you if your LC is fucked just by watching it, you've to test with agar and see, or if you want you can put it in your BRF jars and see if it goes well but it a really risky option.

in my opi nin LC are for expert growers who already knew how to use agar


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OfflineBramscoChill
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Re: liquid culture contamination [Re: mr. whothehell]
    #26492806 - 02/19/20 04:09 AM (3 years, 11 months ago)

I made the LC with an spore syringe indeed (bought from a webshop specialized in spores). The jars seems to grow fine, all white mycelium. My guess is the infection came in with the needle of an syringe when getting the LC out of the jar. I used an glass syringe 2 days ago and some air came into the jar. I thought a glass syringe was more durable and easier to sterilize, bud it's not closing very well.


I just started with cultivating mushrooms, so ha still have to learn allot. I will try testing with agar.

Bud the information I find on the internet is not very consistent. Some say agar isn't the way to detect contermination, some say it does.


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OfflineMrBovineJoni
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Re: liquid culture contamination [Re: BramscoChill] * 1
    #26492811 - 02/19/20 04:23 AM (3 years, 11 months ago)

Quote:

BramscoChill said:
I made the LC with an spore syringe indeed (bought from a webshop specialized in spores). The jars seems to grow fine, all white mycelium. My guess is the infection came in with the needle of an syringe when getting the LC out of the jar. I used an glass syringe 2 days ago and some air came into the jar. I thought a glass syringe was more durable and easier to sterilize, bud it's not closing very well.




Yeah, going from spores to a LC is almost always a crapshoot (very high chance of contamination, and due to the nature of LC - 1% of contamination  == 100% contamination as far as grain inoculation is considered). Very likely that your LC is full of contaminants - the needle here would not change much.

Sorry to say that, but those jars you just posted don't look good at all - almost certainly bacterial and super wet. White does not mean they are not full of contaminants.


Quote:

BramscoChill said:
Bud the information I find on the internet is not very consistent. Some say agar isn't the way to detect contermination, some say it does.



If someone said that agar is not the right way to go to detect contamination in mycology and produce clean cultures - stop listening to anything else they are saying as they could not be more wrong and are misinformed, ignorant or just full of shit.

Agar absolutely is the best way to go in almost every scenario for amateur level mycology.


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OfflineBramscoChill
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Re: liquid culture contamination [Re: MrBovineJoni]
    #26492887 - 02/19/20 06:40 AM (3 years, 11 months ago)

mmm, ok, thanks for the information. That cleared up allot. I will definatly going to try the agar technique. I did not know injection the spores directly in the LC, increases the change on contamination that much...


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OfflineMrBovineJoni
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Re: liquid culture contamination [Re: BramscoChill] * 1
    #26492896 - 02/19/20 06:53 AM (3 years, 11 months ago)

Quote:

BramscoChill said:
mmm, ok, thanks for the information. That cleared up allot. I will definatly going to try the agar technique. I did not know injection the spores directly in the LC, increases the change on contamination that much...



What happens is that there is technically no such thing as clean spores because they grow in open(non-sterile) environment. No one cleans millions of spores under microscope one by one.

So if LC syringe was made from spores (especially by yourself, not a skilled lab technician), it is bound to have some contams in it.


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Offlineshevanel
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Re: liquid culture contamination [Re: MrBovineJoni]
    #26492908 - 02/19/20 07:09 AM (3 years, 11 months ago)

Contermination is cute af


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Offlinemr. whothehell
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Re: liquid culture contamination [Re: BramscoChill]
    #26494280 - 02/20/20 12:37 AM (3 years, 11 months ago)

my sorry to said that they are full of bacteria :frown:

it's always sad to said that t a new cultivator. belive me when you'll learn how to work with agar you'll never turn back.

there's is always the possibility that agar can hide some contams, but the chances to spot them are higher than any other methods that I knew.

where did you by your spores ? I'm form EU too and I'm curios


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OfflineBramscoChill
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Re: liquid culture contamination [Re: mr. whothehell]
    #26496405 - 02/21/20 03:38 AM (3 years, 11 months ago)

I got the spores from magic-mushrooms-shop.com. I am going to try the agar method.

I knew in advance this would happen some time. I threw the infected culture away. I have a look how the jars are doing, so far it is still all white.


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