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Every rye jar contaminated this season
    #25942854 -

So I've started back up and so far I've had every jar of rye get contaminated. Was by no mean an expert before, but I had great success. Using same technique, only difference is I'm using a different brand of rye, gypsum in the soak, and different lid design(ship, not just holes with micropore tape).

I've had to toss my first 7 jars inoculated with MS due to the green goblin. Started 5 more, 1 with MS, and 4 with LC I made from the MS. No growth from the LC jars yet(7 days in), tiny bit from the MS(but already not looking like healthy growth) . I don't know what's changed lol.. I have yet to get healthy myc.

My procedure is: Rinse well, soak no more than 24 hours. Rinse again, boil 30 mins or so, till they are hydrated but have not burst yet. Dry by means of tossing and releasing steam and test with RRs toilet paper test(put rye grain on toilet paper for 15 secs, should be no moisture on it). Load jars, put tinfoil of top and PC for 120 mins. Let. Cool over night. Heat needle till red, cool off with alcohol pad inoc with about 1.5cc. Incubate.

Like I said, I've had success before doing everything the same. I'm trying to figure out what's changed besides and why it would affect my results this way. Any pictures can be taken if need.

Thanks in advance.


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-One Love
Code:
How Frank Gets Shit Done

Edited by Shambosley (04/19/19 04:55 AM)

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Re: Every rye jar contaminated this season [Re: Shambosley]
    #25942949 -

It's more likely to be your spore solution, you should germinate on agar and then take a couple of transfer generations from those, then go to grain..That way you'll be able to clean up any contams that might be in the syringes


--------------------
"How it should & shouldn't look "      "Recognizing contamination"      "Rye Berry Prep"      "Franks G2G"      "Humidity Chamber"      "Agar Recipes"

"How to dial in your monotubs like a Boss"      "Pastywhyte's Easy Agar Tek"      "Elementary [Damion5050]Coir Tek"        "SGFC"      "SAB Tek"      "Easy AF Shoeboxes"


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Re: Every rye jar contaminated this season [Re: Shroomspective]
    #25943674 -

Shroomspective said:
It's more likely to be your spore solution, you should germinate on agar and then take a couple of transfer generations from those, then go to grain..That way you'll be able to clean up any contams that might be in the syringes



Yeah that's kind of the conclusion I have come to, especially considering all my jars have been inoculated with the same MS    syringe.
Never done agar, I've always been intimidated by it but it might be time to give it a try


--------------------
-One Love
Code:
How Frank Gets Shit Done

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Re: Every rye jar contaminated this season [Re: Shambosley]
    #25943766 -

Agar is easy..you've got the main thing being a PC, just follow the easy agar tek in my sig, and there's some agar recipes too to get you started


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"How it should & shouldn't look "      "Recognizing contamination"      "Rye Berry Prep"      "Franks G2G"      "Humidity Chamber"      "Agar Recipes"

"How to dial in your monotubs like a Boss"      "Pastywhyte's Easy Agar Tek"      "Elementary [Damion5050]Coir Tek"        "SGFC"      "SAB Tek"      "Easy AF Shoeboxes"


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Re: Every rye jar contaminated this season [Re: Shroomspective]
    #25945642 -

Shroomspective said:
Agar is easy..you've got the main thing being a PC, just follow the easy agar tek in my sig, and there's some agar recipes too to get you started


The pastywhytes easy agar tek link says the recipe is potato flakes, agar,  and Karo or honey. But in the agar recipes link, the "potato flakes" along with all the others call for the use of yeast. Why is that? Is it possible to substitute yeast with Karo or must I have yeast for any agar recipe?


--------------------
-One Love
Code:
How Frank Gets Shit Done

Extras: Filter Print Post Top
Re: Every rye jar contaminated this season [Re: Shambosley]
    #25945691 -

Agar can be PDA or PDYA, you don't have to use yeast, but I add it as it provides some extra nutrients, and in my grows its always proven helpful


--------------------
"How it should & shouldn't look "      "Recognizing contamination"      "Rye Berry Prep"      "Franks G2G"      "Humidity Chamber"      "Agar Recipes"

"How to dial in your monotubs like a Boss"      "Pastywhyte's Easy Agar Tek"      "Elementary [Damion5050]Coir Tek"        "SGFC"      "SAB Tek"      "Easy AF Shoeboxes"


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Re: Every rye jar contaminated this season [Re: Shroomspective]
    #25982063 -

So I'm dabbling in agar, used the Bods mea agar pour recipe(10g agar, 10g extra light malt extract, 1g yeast) And everything is clean an sterile before inoculating with Ms syringe. I tried streaking plate the best I could. Of course it being my first time I got a little carried away as I didn't do the zig zag, I spread one drop all around with inoculation loop.
I know inoculating with spore syringe is the worst but it was kind of my only option as previously mentioning my syringe has to be contaminated. Anyways so I did 5 plates to try and get something viable. So far I only got growth on one and there are two different visible growths.
Here's a picture, what do I got going on here lol



--------------------
-One Love
Code:
How Frank Gets Shit Done

Edited by Shambosley (05/09/19 09:09 PM)

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Re: Every rye jar contaminated this season [Re: Shambosley]
    #25983238 -

Bright white spot looks somewhat like germinated mycelium (probably) the rest is mold..how long has this been going?

Try take some transfers from within the bright white patch to new plates and see what happens


--------------------
"How it should & shouldn't look "      "Recognizing contamination"      "Rye Berry Prep"      "Franks G2G"      "Humidity Chamber"      "Agar Recipes"

"How to dial in your monotubs like a Boss"      "Pastywhyte's Easy Agar Tek"      "Elementary [Damion5050]Coir Tek"        "SGFC"      "SAB Tek"      "Easy AF Shoeboxes"


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Re: Every rye jar contaminated this season [Re: Shroomspective]
    #25984158 -

Started five plates, 10 days ago. Picture was taken on day 8, first sign of growth was on about day 5 with just the bright white spot. The fuzzy stuff came up on about day 7 and grew really fast, Which had me concerned that it definitely wasn't mycelium. I just thought if the plate had bacteria it would show up faster than that. I'm having terrible luck with this ms and agar. MS on agar is hard lol

I do have about 4 or 5 quart jars that should reach 100% colonization within the next couple days(not as many as I wanted), but I might spawn them and try to get a fruit body to clone on agar.


--------------------
-One Love
Code:
How Frank Gets Shit Done

Extras: Filter Print Post Top
Re: Every rye jar contaminated this season [Re: Shambosley]
    #25984172 -

If those 4-5 jars are 100% clean i.e no wet spot etc, then why not use one to go G2G, you can usually do 1:5 per jar, so you could make another 5 and then have 4+5 to spawn, plus take some colonised grains and put them on agar to get your plates on track, then you have a viable ongoing supply to use


--------------------
"How it should & shouldn't look "      "Recognizing contamination"      "Rye Berry Prep"      "Franks G2G"      "Humidity Chamber"      "Agar Recipes"

"How to dial in your monotubs like a Boss"      "Pastywhyte's Easy Agar Tek"      "Elementary [Damion5050]Coir Tek"        "SGFC"      "SAB Tek"      "Easy AF Shoeboxes"


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Re: Every rye jar contaminated this season [Re: Shroomspective]
    #25986145 -

That's a good idea and I just may do that.  With my luck lately I thought I might just go with what I know and that's spawning grain to bulk lol so far agar has gotten my spirit down and I've never done G2G, guess I always did it the harder way before. Ms-grain-spawn2bulk-clone w/ LC then Lc-grain lol

How do I tell if my agar is being contaminated during the pour, like how long would a non inoculated agar start to grow bacteria if contaminated.


--------------------
-One Love
Code:
How Frank Gets Shit Done

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Re: Every rye jar contaminated this season [Re: Shambosley]
    #25986201 -

Mine have had issues around 5 days into incubation with or without mycelium. Pouring agar hot is great for contamination control, I leave fresh poured plates in an airtight lunchbox (sprayed with isopropyl alcohol) to cool down with no air movement. With this box I very rarely get contams. Big fan of agar>grain>bulk

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Re: Every rye jar contaminated this season [Re: mojotoke]
    #26006784 -

mojotoke said:
Mine have had issues around 5 days into incubation with or without mycelium. Pouring agar hot is great for contamination control, I leave fresh poured plates in an airtight lunchbox (sprayed with isopropyl alcohol) to cool down with no air movement. With this box I very rarely get contams. Big fan of agar>grain>bulk


Yeah I've noticed a few of mine are getting contams just from sitting. I only have them back in the bag the plates came in though. Id say about 20% of them maybe I'll try a better storing alternative.
Do you do agar wedge to grain? I've had no luck with agar  so far but Ive got a little something going on. I G2G'd one of the jars I had as @shroomspective had mentioned and got 7 more going. I'm real impressed with G2G as this was my first time.
While I was at it, I put a a couple remaining grains to agar, and seem to have some growth. Since I've never had healthy mycelium growth on agar I'll post a picture soon for maybe someone experienced to help me to know if it's actually healthy. TIA.


--------------------
-One Love
Code:
How Frank Gets Shit Done

Extras: Filter Print Post Top
Re: Every rye jar contaminated this season [Re: Shambosley]
    #26007037 -

I'm no expert but the last pic you posted didn't look very healthy, the light patch looks like a contaminant &the  two white dots like agitated mycelium. You could wait for the full plate to grow though and try cutting out healthy "rhizomorph" and/or "tomentose" growth. These pure mycelium pieces can be used to start new plates that hopefully aren't contaminated.

Yeah I do wedge to grain all the time, supplementing grain with agricultural gypsum and bran (wheat) speeds up colonisation so much and increases yield a bit, highly recommend supplementing spawn if you have very sterile procedure and can seal your bags

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Re: Every rye jar contaminated this season [Re: mojotoke]
    #26008026 -

mojotoke said:
I'm no expert but the last pic you posted didn't look very healthy, the light patch looks like a contaminant &the  two white dots like agitated mycelium. You could wait for the full plate to grow though and try cutting out healthy "rhizomorph" and/or "tomentose" growth. These pure mycelium pieces can be used to start new plates that hopefully aren't contaminated.


Most definitely was contaminated, tossed it along with all my other onse. Didn't get any to grow from MS to agar. What is that brownish stuff I'm sessing, seems to be everywhere my spore solution went? When I did my grain to agar it had the same thing wherever the grain rolled around on the agar.


--------------------
-One Love
Code:
How Frank Gets Shit Done

Extras: Filter Print Post Top
Re: Every rye jar contaminated this season [Re: Shambosley]
    #26008461 -

Yeah grain to agar can be really tricky and I've never had success with it. Even with syringes and tissue samples I get a high contamination rate. Agar is tricky without a SAB or a flow hood but you can just do so much with a good plate

This 'tinfoil' you put on your jars before PC is most probably the problem. You should use the original jar lids that seal shut, poke/drill some holes in the lids, cover these holes with microporous tape for air filtration, TBEN add the tinfoil if you want.

If you're doing tinfoil with no jar lid the grain contaminate very easily.

LC's can get contaminated but it's uncommon

Please though, store your agar in an airtight, air filtered box. Unfiltered moving air is by far the biggest threat to any grow

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Re: Every rye jar contaminated this season [Re: mojotoke]
    #26008645 -

mojotoke said:
If you're doing tinfoil with no jar lid the grain contaminate very easily.


Im not really having grain contamination trouble anymore. I finally got a few clean jars from some nasty syringes I guess, and G2Gd one jar to make 7 more, and spawned the other 4.

I'm now working on agar and trying to get some clean strong mycelium, maybe I need to make a new post for this journey. So far all I've got is 3 plates inoculated with grains and I'm seeing some decent growth. Even if there are some contaminates, I'm under the impression with agar, I can transfer clean mycelium away from the contams.



mojotoke said:
Please though, store your agar in an airtight, air filtered box. Unfiltered moving air is by far the biggest threat to any grow


Yes! I definitely need an air tight container for storing my agar plates. Right now my inoculated plates are just incubating with my jars in a normal tub and my clean onse are just in the bag the plates came in.


--------------------
-One Love
Code:
How Frank Gets Shit Done

Extras: Filter Print Post Top
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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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