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Offlinegreggor22
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How long till Agar plates show contams?
    #14137337 - 03/17/11 03:26 PM (12 years, 10 months ago)

I did some searching and saw suggestions to do various things to check your lc for contam, brf cakes and agar plates were the 2 most prevelent and I don't really feel like making up a batch of cakes right now so I did agar which I had available what I didn't see was a definitive answer to how long I should wait for contams to show themselves?

I am trying to check some LCs, I poured agar plates, kept a couple as control and none show contam.  did about 10 tests and some are showing cobweb and one is showing trich.  Its only been like 3 days.  How long till it is safe to consider the LC clean?  is 1 week sufficient or should I wait longer?

PS the lcs I am testing are 3 months old thats why such a high contam rate.  I had no intention of using these but I wanted to use them for practice on my next batch of LCs since which are currently cooking, I'm looking to knock up like 30 jars and I don't want to take any chances.


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Offlinegreggor22
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Re: How long till Agar plates show contams? [Re: greggor22]
    #14148258 - 03/19/11 12:40 PM (12 years, 10 months ago)

anyone have a straight answer?


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InvisibleDoc_T
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Re: How long till Agar plates show contams? [Re: greggor22]
    #14148291 - 03/19/11 12:49 PM (12 years, 10 months ago)

Two different answers- an agar dish should show contams in 2 or three days, and certainly by a week.

But not every contaminated LC is going to show anything, you can have what looks like normal myc but isn't.
Until you've taken jars all the way to fruiting, you can't be certain your LC is capable of producing fruits.

I'd use the agar to clean up a bit of myc, and just toss the LC. If it was me.


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Offlinegreggor22
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Re: How long till Agar plates show contams? [Re: Doc_T]
    #14148640 - 03/19/11 02:16 PM (12 years, 10 months ago)

thanks for taking the time to answer Doc.  On the scale I'm doing Agar wedges would be untenable as an innoc method.  I don't feel like plating 30 dishes of agar to innoc 100 jars (edibles btw)

Not to mention I'm not that good with a glovebox yet, so anything I can do to avoid opening an uncolonized jar is good for me.  LC has always worked well for me, I rarely have bad batches and I just wanted to start taking that extra step to hopefully increase my sucess rate a little.


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OfflineAjaxx
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Re: How long till Agar plates show contams? [Re: greggor22]
    #14150564 - 03/19/11 08:38 PM (12 years, 10 months ago)

fuckin A

is there any point to use agar? i just ordered a kit so i could make some agar to test my LCs... did i just waste a bunch of money or what?


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InvisibleDoc_T
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Re: How long till Agar plates show contams? [Re: Ajaxx]
    #14150576 - 03/19/11 08:41 PM (12 years, 10 months ago)

Yes, but not the way you think.
You wasted money on LC.
And on buying an agar kit, vs shopping locally.

But agar is gold.

To address the point about doing 100 jars, you don't start those with agar wedges, you do g2g to get there.


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OfflineAjaxx
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Re: How long till Agar plates show contams? [Re: Doc_T]
    #14150609 - 03/19/11 08:46 PM (12 years, 10 months ago)

Quote:

Doc_T said:
Yes, but not the way you think.
You wasted money on LC.
And on buying an agar kit, vs shopping locally.

But agar is gold.

To address the point about doing 100 jars, you don't start those with agar wedges, you do g2g to get there.




well i guess i didnt waste any money making the LCs.

i cant get agar or anything that comes in the kit anywhere in town, so i ordered online. :shrug:

guess ill play around with the agar and see how it goes.


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Re: How long till Agar plates show contams? [Re: Ajaxx]
    #14152503 - 03/20/11 06:27 AM (12 years, 10 months ago)

Quote:

i cant get agar or anything that comes in the kit anywhere in town,




I call BS. Where do you live?
And what comes in the kit besides agar?


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Offlinequebus
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Re: How long till Agar plates show contams? [Re: Doc_T]
    #14153180 - 03/20/11 11:03 AM (12 years, 10 months ago)

I tried all the local stores from Shaws to Walmart to Stop & Shop.  Nada.  I even went to the local asian food market - they never heard of it.  I ordered agar flakes online.  We used to have a hobby store but they are closed now.


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OfflineAjaxx
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Re: How long till Agar plates show contams? [Re: Doc_T]
    #14153351 - 03/20/11 11:37 AM (12 years, 10 months ago)

Quote:

Doc_T said:
Quote:

i cant get agar or anything that comes in the kit anywhere in town,




I call BS. Where do you live?
And what comes in the kit besides agar?




http://everythingmushrooms.com/shop/Agar-Culturing-Kit-Advanced.html


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OfflineTerry M
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Re: How long till Agar plates show contams? [Re: Ajaxx]
    #14153487 - 03/20/11 12:11 PM (12 years, 10 months ago)

I've made up a bunch of LC jars, and have been using them. They seemed great at first -- tons of inoculations compared to a spore or LC syringe. But now I see that they store in the fridge no longer than syringes, take up a lot of space, and boy, do they hide contamination! I am rapidly becoming a fan of plates! :wink: I may be selling/trading some nice, homemade LC lids with self-sealing injection ports and Whatman type filters real soon!


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Invisiblebiologys
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Re: How long till Agar plates show contams? [Re: Terry M]
    #14153494 - 03/20/11 12:12 PM (12 years, 10 months ago)

you're telling me a local asian market said they never heard of agar-agar?


that makes absolutely no sense!


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OfflineTerry M
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Re: How long till Agar plates show contams? [Re: biologys]
    #14153527 - 03/20/11 12:20 PM (12 years, 10 months ago)

I buy premixed agar from Fungi Perfecti, and am very happy with it. Plus, they sell antibacterial agar for doing agar sandwich isolation. For the latter, I found nowhere to buy gentamicin in small quantities.


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Offlinegreggor22
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Re: How long till Agar plates show contams? [Re: Terry M]
    #14156632 - 03/20/11 10:08 PM (12 years, 10 months ago)

Doc, again thanks for the answer, and also the advice, g2g is not someting I find fun so I'm gonna pass on that.  Like I said I'm not that good with a glovebox.  I'm sure I'm missing out, and working on those skills is definately in my list of things to do, but for now my LCs do work for me very well.  Just wanted to do a little pre screening with the agar.

As far as asian store not knowing what Agar agar was, I'm not that surprised, the asian store I shop has 1 person who can speak any english, and its very broken at best lol.  i'm willing to guess they have it, but finding something in those stores is a royal PITA if you haven't shopped them dozens of times or don't know the language... I actually asked the asian store to help me find it she claimed she had no idea what it was, then after browsing a bit more I did find it, lol it was actually hung on the side of their coke fridge beside the cash register... Big asian lettering, tiny english, but I did find it eventually.  Sometimes Its helpful to find a customer who looks like they speak english to help translate.


and btw testing your LC's on Antibacterial agar is not a good call, you are using a product designed to stop contams from growing to check if you have a contam...


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Offlineafrosheen
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Re: How long till Agar plates show contams? [Re: greggor22]
    #14156951 - 03/20/11 10:58 PM (12 years, 10 months ago)

For the agar agar, I wouldn't count on the people there knowing what or where it is. It's usually in the spice aisle next to all the other strange ingredients you never knew existed (and even gypsum for cooking). I swear you could make fireworks with the stuff they have there.

At any rate, as a real control, you should have taken the lids off of your control dishes in the glovebox when you were inoculating your test dishes. That'd test the sterility of your GB and your technique. I'm not saying you'd have every lid off for the duration, but each time you removed a lid from a test dish you'd remove the lid from the control also, then place them back on in sequence.

Cobweb and trich are both floating around in the air everywhere, so it's hard to say if your agar dishes were contaminated from the LC or during inoculation in the GB. My first experience with agar I transferred wild prints to plates and got some clean, some dirty, and one that was 100% cobweb and nothing else. However the second transfer to new plates had zero contams so I knew my GB technique was acceptable.

I agree with Doc on going from agar to grains. Take a half-full quart with sterile grains or a half pint, drop an agar wedge in, sit back and wait. Eventually you can g2g to full sized grain jars and go from there, the whole 1-10-100 thing.

>Edit:
Quote:

  Like I said I'm not that good with a glovebox.


Man you might as well give up now or build a flowhood. Glovebox technique is critical for many aspects of this hobby. If you can't master a glovebox you will always have contams regardless of what you're trying to accomplish.


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Edited by afrosheen (03/20/11 11:00 PM)


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InvisibleDoc_T
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Re: How long till Agar plates show contams? [Re: quebus]
    #14158112 - 03/21/11 06:30 AM (12 years, 10 months ago)

Quote:

phortre55 said:
I tried all the local stores from Shaws to Walmart to Stop & Shop.




:facepalm:

I don't know if you're trolling but I'm done with this thread.


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You make it all possible. Doesn't it feel good?


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