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OfflineRooster Cogburn
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Let's see what is inside of coir * 1
    #23542736 - 08/15/16 01:30 AM (7 years, 5 months ago)

This is an open experiment, I encourage anyone to try

I was having a conversation with Pasty in another thread about coir, specifically about how much life is actually in coir when we get it. The consensus seems to be that it's pretty damn clean and even heat treated before  we get it. I'm skeptical about that assumption, however I do know for a FACT that coir performs well any way that it is prepared, be it just hydrated, pasteurized and treated with boiling water. It seems also the general consensus is that coir is so clean it is actually sterilized with the bucket method, and so depraved of nutrients that competitor molds and bacteria can't get a foot on it.
Pasty said he wanted to do an experiment about the initial load there actually is in coir when we get it to put the subject to rest, so I was making agar tonight and thought what the hell, I will start one as well.  Let's see if there is anything to germinate.

For this experiment I am using MEA


The coir being used is this, botanicare coir, I have used it for years and never had an issue with it, it's great and never has wierd debris.


This experiment will use samples from 4 different forms of coir:
1. Dry coir
2. Hydrated untreated coir
3. Pasteurized coir
4. Coir treated with boiling water

Here are the samples, the bag is pasteurized CVG



I am placing samples of each on dishes of agar and incubating them. I did this with a scalpel scooping the samples onto the dish and pushing it into the agar a bit.


Anyways, lets see how much shit is in there if you actually try to grow it and find out just why coir is the most efficient and forgiving substrate for cubensis cultivation.


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Offlinelukehighwalker710
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Re: Let's see what is inside of coir [Re: Rooster Cogburn]
    #23542747 - 08/15/16 01:38 AM (7 years, 5 months ago)

:popcorn:
I suggest adding PC sterilized coir to the list, and probably better to pastuerize some coir yourself for the experiment rather than pre-pastuerized vendor bags. Though, those could very well be another one on the list.

Either way, I'm on board to see how this turns out.


--------------------
:vaped: Highwalker :bongload:


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InvisiblebodhisattaMDiscordReddit
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Re: Let's see what is inside of coir [Re: Rooster Cogburn]
    #23542749 - 08/15/16 01:40 AM (7 years, 5 months ago)

coir pith presses exert an enormous amount of pressure on the fiber that is mixed with a binding agent. the heat is produced from the pressing. compression = heat. it's how a diesel engine works, not to mention the friction forces from all those fibers getting pressed into such a small brick.

mycelium is able to fend for itself pretty well after the spawn step. the sterilization is only to remove all vegetative life. spores and endospores can survive if they're around but it's still called sterilization.

MEA is ok, for this test,

what you really should do is make sterile water of a known volume, and then add a coir chunk from the inside of the brick, have weights of the jar with the sterile water and then weigh it after adding the coir, then you'll know how much the coir weighed you added without weighing it and contaminating it.

then let it dissociate into the water.

take 1CC of homogeneous coir water and then do a pour plate and a spread plate.

you should use plate count agar.

you should have a control plate that you do the same techniques with just plain sterile water and carry out the exact same maneuvers.

ideally you would do the experiment twice and incubate the other half of the plates anaerobically

it's a good place to start, what you're doing. if you get any positives it would be grounds for a real investigation


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OfflineRooster Cogburn
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Re: Let's see what is inside of coir [Re: lukehighwalker710]
    #23542754 - 08/15/16 01:41 AM (7 years, 5 months ago)

That's my pasteurized coir, that is my preferred method of doing it personally.
And your right I should add pc sterilized on there:thumbup:


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InvisibleHashed
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Re: Let's see what is inside of coir [Re: Rooster Cogburn]
    #23542758 - 08/15/16 01:45 AM (7 years, 5 months ago)

:threadmonitor:


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OfflineRooster Cogburn
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Re: Let's see what is inside of coir [Re: bodhisatta]
    #23542762 - 08/15/16 01:47 AM (7 years, 5 months ago)

Quote:

Trusted Cultivator said:
coir pith presses exert an enormous amount of pressure on the fiber that is mixed with a binding agent. the heat is produced from the pressing. compression = heat. it's how a diesel engine works, not to mention the friction forces from all those fibers getting pressed into such a small brick.

mycelium is able to fend for itself pretty well after the spawn step. the sterilization is only to remove all vegetative life. spores and endospores can survive if they're around but it's still called sterilization.

MEA is ok, for this test,

what you really should do is make sterile water of a known volume, and then add a coir chunk from the inside of the brick, have weights of the jar with the sterile water and then weigh it after adding the coir, then you'll know how much the coir weighed you added without weighing it and contaminating it.

then let it dissociate into the water.

take 1CC of homogeneous coir water and then do a pour plate and a spread plate.

you should use plate count agar.

you should have a control plate that you do the same techniques with just plain sterile water and carry out the exact same maneuvers.

ideally you would do the experiment twice and incubate the other half of the plates anaerobically

it's a good place to start, what you're doing. if you get any positives it would be grounds for a real investigation




:thumbup:
Well I'm glad I posted this so we can discuss some different methods of going about this, you have some good ideas there.


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InvisiblebodhisattaMDiscordReddit
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Re: Let's see what is inside of coir [Re: Rooster Cogburn] * 1
    #23542767 - 08/15/16 01:53 AM (7 years, 5 months ago)

then you can enumerate CFU/g of coir

say you use 100mL of water in a jar and sterilize it then weigh it and it weighs 300 grams we assume 100g is water. then add coir. you weigh it after adding coir and it's 310g. so we know 10g of coir in 100mL water.

we plate 1mL of coir water in a pour plate.

then if you get any colonies in the plate since you did a pour plate you can count the colonies.

then say you counted 10 colonies. you would have 10cfu per 1ml and 1ml has .1g coir since there's 10g in 100mL

so then you have a CFU count of 100 CFU/1g coir

then any one else who tests it can compare the numbers


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Re: Let's see what is inside of coir [Re: bodhisatta]
    #23542779 - 08/15/16 02:04 AM (7 years, 5 months ago)



these are pour plates of a badly infected beer. they're plated anaerobic since beer is obviously anaerobic. and the only spoilers of beer are hence anaerobic.

this was 1mL of finished bottled beer. there's too many colonies to count. it's easily over 500. let's just say 1000 CFU per mL

which means that 3785mL in a gal so there's at least 3.785 million bacterium in every gal. delicious. 354,800 in every 12 oz pour.

needless to say the QA plate was just for fun the beer was too gross to sip.

what you do though is pipette the 1mL sample into an empty dish then pour 47C agar over the top, just hot enough to still be liquid but not hot enough to kill any of the microorganisms. swirl the plate gently and let it harden. this way you get colonies forming in the agar as pinpoints and you can count them.


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OfflineRooster Cogburn
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Re: Let's see what is inside of coir [Re: bodhisatta]
    #23542834 - 08/15/16 02:54 AM (7 years, 5 months ago)

Dude, this is great info. I will definitely try this, sounds fun too.
I just used the last of my MEA, what would you suggest a good recipe for this type of work if I was to go pick up ingredients tomorrow instead of ordering more premixed?


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Offlineinvitro


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Re: Let's see what is inside of coir [Re: Rooster Cogburn]
    #23543035 - 08/15/16 06:34 AM (7 years, 5 months ago)

:popcorn:


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Re: Let's see what is inside of coir [Re: invitro]
    #23543432 - 08/15/16 10:04 AM (7 years, 5 months ago)

0.5% peptone
0.25% yeast extract
0.1% glucose
1.5% agar
pH adjusted to neutral at 25 C.

That's plate counts recipe. Malt extract probably does plenty fine for us tho


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OfflineRooster Cogburn
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Re: Let's see what is inside of coir [Re: bodhisatta]
    #23544149 - 08/15/16 01:45 PM (7 years, 5 months ago)

Thanks, I have only used MEA since I started growing, I don't know why, just creature of habit and plus it's the main formula you can buy premixed.
It will actually be pretty fun using a new recipe and seeing how my cultures react to it after being fed and stored on MEA for so long.


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Offlineblackout
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Re: Let's see what is inside of coir [Re: Rooster Cogburn]
    #23544227 - 08/15/16 02:09 PM (7 years, 5 months ago)

Worthwhile experiment :thumbup:

Quote:

Rooster Cogburn said:
And your right I should add pc sterilized on there:thumbup:



I thought the same at first, but then thought why bother, nothing will grow, this will be of no surprise or interest to anybody. Microwaving is another method people would have doubts about. I would also be interested in chemical sterilization methods like using peroxide or bleach, but this is not commonly done.

An experiment I would like to see is adding colonised grain from an isolate to coir treated in different ways and see how it performs. The problem with this is having them all equal hydration wise. I usually measure hydration levels by weighing after soaking stuff. However most people seem to find that after being heat treated that coir can take on more moisture.

I wonder if there is some way to try and get equal levels of "field capacity", rather than equal water hydration (which would be grams water per gram of dry coir). Maybe if the coir was put in a coffee french press and a weight put on top and pour off the liquid.

If actually growing with grains & coir I would be interested in seeing how the PC'd coir performs.


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Re: Let's see what is inside of coir [Re: blackout]
    #23544238 - 08/15/16 02:14 PM (7 years, 5 months ago)

PC sterilized acts as a control. If you get positive with that maybe your got a flaw somewhere


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Re: Let's see what is inside of coir [Re: bodhisatta]
    #23544923 - 08/15/16 06:20 PM (7 years, 5 months ago)

Well this saves me some trouble. Great stuff.


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OfflineRooster Cogburn
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Re: Let's see what is inside of coir [Re: Pastywhyte]
    #23544956 - 08/15/16 06:30 PM (7 years, 5 months ago)

I figure you have enough on your plate, ooh pun.


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OfflineRooster Cogburn
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Re: Let's see what is inside of coir [Re: Rooster Cogburn]
    #23545463 - 08/15/16 09:40 PM (7 years, 5 months ago)

Ok guys, I have some action going on tonight in the bacteria department.

Let's look at these samples



First, here is the dry coir



And the hydrated




Treated with boiling water


And Pasteurized



Here are my control plates, and like you guys are saying pc sterilized coir would be a great control along with this.



:thumbup:


Edited by Rooster Cogburn (08/15/16 09:42 PM)


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Re: Let's see what is inside of coir [Re: Rooster Cogburn] * 1
    #23545482 - 08/15/16 09:47 PM (7 years, 5 months ago)

Not bad. I wouldn't have suspected the dry to have as much as it does but it's really not even that bad really. No mold yet either.


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OfflineRooster Cogburn
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Re: Let's see what is inside of coir [Re: Pastywhyte]
    #23545515 - 08/15/16 09:58 PM (7 years, 5 months ago)

Yeah it's not bad at all, I expect tomorrow to be mold day if it's there or wants to germinate.


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Re: Let's see what is inside of coir [Re: Rooster Cogburn]
    #23545553 - 08/15/16 10:10 PM (7 years, 5 months ago)

To be honest I would have expected a few more phenotypes to be present/obvious. I would love for bodhi to get that under a scope. Bet it's an endospore forming species.


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