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Offlinejackrabbitcytoplas
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Re: Please help me get this right, contamination blues.. [Re: Anastomosis]
    #26852800 - 07/29/20 11:30 PM (3 years, 5 months ago)

Did you consider trying muda bottles?
Eliminating a few stages could help localizing the source of contaminations.

Just throwing it out there, sorry if I'm missing something obvious.


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: jackrabbitcytoplas]
    #26852803 - 07/29/20 11:32 PM (3 years, 5 months ago)

Ya know I never tried muda bottles. I thought the loosened lid as air exchange was shady so I never bothered trying..


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26852804 - 07/29/20 11:34 PM (3 years, 5 months ago)

Didn't seem to be an efficient means to grow a bunch of mushies either. I mean maybe it is and I'm mistaken, but my girlfriend eats a lot of mushrooms :lol:


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


Edited by Sherlock Shrooms (07/29/20 11:35 PM)


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26852805 - 07/29/20 11:35 PM (3 years, 5 months ago)

Oh I see, as a means of identifying the source of contamination. How so?


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


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Offlinejackrabbitcytoplas
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26852813 - 07/29/20 11:46 PM (3 years, 5 months ago)

Quote:

Sherlock Shrooms said:
Oh I see, as a means of identifying the source of contamination. How so?




Because you're sterilizing the whole thing, inoculating once from a clean plate (via LI) and then the next time you're exposing it to open air is after the mycelium colonized the bottle.
You're not expanding G2G, you're not spawning and you don't have it colonizing in a non sterile container supposedly exposed to your spore load.

If bottles comes out 100%, it could mean one of the things you didn't have to do for bottles is the problem.


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InvisibleAnastomosis
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26852844 - 07/30/20 12:17 AM (3 years, 5 months ago)

Quote:

Sherlock Shrooms said:
Quote:

Anastomosis said:
Quote:

Anastomosis said:
If some tubs are good and some are not, the problem could be your technique.






For sure. I'll definitely be paying closer attention with my sterile work. I appreciate the advice, and it almost fits. However I am a bit skeptical because I haven't seen a single jar or plate contam in well over a year, no BS not a single one. If my technique was bad, as it was when I started growing, then I'd see an occasional jar or plate get contam'd like I used to. No one's perfect but I'm damn careful, and if my technique was so bad that I was losing half of my tubs then I would definitely be seeing some contams in plates or jars sometimes. I can't even remember the last time I saw a plate or jar contam... I treat sterile work like I'm handling biological warfare that would wipe out all of humanity if I slip.



Your technique for how you handle plates vs a2g vs g2g are all different. It's the g2g I'm talking about.


--------------------
"120mph with a trailer full of dogs and a truck full of drug addicts."


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Invisiblesandman420
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26852956 - 07/30/20 04:31 AM (3 years, 5 months ago)

Quote:

Sherlock Shrooms said:
I'm using a flowhood I built. This

I make agar with 7.5 grams of malt extract and 10 grams of agar powder per 500mL of water. I do A2G into rye berries that have been soaked 6 to 8 hours, brought to a simmer until the middle is fully gelatinized, and then strained and spread out on drying racks until the grains are dry to the touch with a little bit of a whitish look to the surface before I load them into jars. Then I PC for 2 hours and let cool overnight and load directly into my flowhood from the PC the next day. I flame sterilize with a map gas propane torch that sits right at the edge of my flowhood. I wipe down the entire flow hood with alcohol and anything that gets brought inside of it then let it run for a while before I start doing transfers. I do all my work about a few inches in front of the filter..




You need to lift that rack up several more inches. The bottom of a flowhood is full of turbulence that brings a backwash of contaminated air alllllllllll the way up on the bottom.

i can show you with a particle counter how bad the bottom of a flowhood is.

Never work on plates that close to the bottom anyone reading this.

When you first described your problems to me it sounded like you have trichaderma/penicillium/whatever green mold that has gone myco-parasite on your mycelium. I don't know what cubesnsis mycelium looks like when this has occurred. No one probably does because you need a scanning electron microscope to see the interaction. Here is an interesting article on similar

The interaction pics are towards the bottom.

But your plates etc don't really look very off or anything, for the most part at least.

Lift that rack up.


--------------------
- Sandbag Tek - How To Sterilize Spawn Bags - All About Static Pressure / Pressure Drop for DIY Flow Hoods - Sandman's LC Tek-

Marijuanaut escapes earth to cultivate - Grow-room is church temple of the new stoner breed


Edited by sandman420 (07/30/20 04:40 AM)


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: jackrabbitcytoplas]
    #26853044 - 07/30/20 06:53 AM (3 years, 5 months ago)

Quote:

jackrabbitcytoplas said:
Quote:

Sherlock Shrooms said:
Oh I see, as a means of identifying the source of contamination. How so?




Because you're sterilizing the whole thing, inoculating once from a clean plate (via LI) and then the next time you're exposing it to open air is after the mycelium colonized the bottle.
You're not expanding G2G, you're not spawning and you don't have it colonizing in a non sterile container supposedly exposed to your spore load.

If bottles comes out 100%, it could mean one of the things you didn't have to do for bottles is the problem.



That's a good idea, the quesion I have is that it's kind of like fruiting out of a jar and just because it doesn't contam how would I know that it's not the fact that it didn't have to recolonize in open air? I've always thought that was the archilles heel of our common cube method, the 5 or 6 days it takes to colonize a substrate once exposed to open air/pandoras box of contaminants..? Like if a muda bottle goes well compared to the tubs I've been spawning, does that mean it wasn't exposed to the sporeload in my home while recolonizing or does it mean I didn't introduce a contam during my typical grain to grain process? I feel like I'd still be at square one where I am right now, because those seem to be the 2 likely possibilities.

And then again if it is my grain prep, whether it's the burst grains or endospores germinating in the spawn again, those things cause problems once the grain spawn is broken up in to a tub and recolonizing a substrate. If it never recolonizes I can almost guarantee it won't contam. Then I'm still in the situation I am currently in. And if it does contam I'm still in the same situation. Maybe I'm missing something feel free to correct me. I'd just hate to go through an entire process where either result doesn't tell me anything...

And several months ago I had a dozen or so spawn jars of 2 different varieties that sat around for a few weeks fully colonized so I decided to just case them and fruit them out of the top of the jar. Every single one of them didn't contam for like 3 or more flushes until I threw them out.


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: sandman420]
    #26853050 - 07/30/20 06:57 AM (3 years, 5 months ago)

Quote:

sandman420 said:
Quote:

Sherlock Shrooms said:
I'm using a flowhood I built. This

I make agar with 7.5 grams of malt extract and 10 grams of agar powder per 500mL of water. I do A2G into rye berries that have been soaked 6 to 8 hours, brought to a simmer until the middle is fully gelatinized, and then strained and spread out on drying racks until the grains are dry to the touch with a little bit of a whitish look to the surface before I load them into jars. Then I PC for 2 hours and let cool overnight and load directly into my flowhood from the PC the next day. I flame sterilize with a map gas propane torch that sits right at the edge of my flowhood. I wipe down the entire flow hood with alcohol and anything that gets brought inside of it then let it run for a while before I start doing transfers. I do all my work about a few inches in front of the filter..




You need to lift that rack up several more inches. The bottom of a flowhood is full of turbulence that brings a backwash of contaminated air alllllllllll the way up on the bottom.

i can show you with a particle counter how bad the bottom of a flowhood is.

Never work on plates that close to the bottom anyone reading this.

When you first described your problems to me it sounded like you have trichaderma/penicillium/whatever green mold that has gone myco-parasite on your mycelium. I don't know what cubesnsis mycelium looks like when this has occurred. No one probably does because you need a scanning electron microscope to see the interaction. Here is an interesting article on similar

The interaction pics are towards the bottom.

But your plates etc don't really look very off or anything, for the most part at least.

Lift that rack up.



Holy crap that makes sense! I'll lift the rack up and see if that helps. Again it's weird though that for over a year I haven't seen a single contam in a plate or jar. I'd think if I was occasionally wafting random spores into my plates then surely at some point I'd see a contam in my work before it goes to tubs. But either way I'll lift up the rack to eliminate the variable. Good tip!


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26853054 - 07/30/20 07:00 AM (3 years, 5 months ago)

Right now I've got all my spawn created for this round so all these tips with my grain prep, raising the rack on my flowhood, fruiting from jars or creating muda bottles, and sharpening up my grain to grain transfers will take some time to get feedback on. Once I go through those steps I'll log them all here and let you guys know how it goes. So far I've gotten several good insights. Gratitude


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


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Invisiblerickyswamps
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26853057 - 07/30/20 07:02 AM (3 years, 5 months ago)

You are overthinking your grain prep for sure.  They don't need to be soaked or rinsed, just boiled and hydrated.  A few burst grains are no big deal.  What kind of bags are you using (filter size) and how are they sealed?  What is your water source for spawning?

Your pics aren't particularly helpful, because the plate looks good, the jar looks good, the bag looks good I think, one tub looks good, but the other doesn't.  But they aren't even connected to each other.  We would need to see all the jars or bag that was used to spawn that green tub.  Also, was it a proven variety that you had success with before or something new?  Sometimes I carry from plate to tub a new variety and it goes horribly.  I think the plate is good, but I'm wrong...


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: rickyswamps]
    #26853083 - 07/30/20 07:19 AM (3 years, 5 months ago)

Quote:

rickyswamps said:
You are overthinking your grain prep for sure.  They don't need to be soaked or rinsed, just boiled and hydrated.  A few burst grains are no big deal.  What kind of bags are you using (filter size) and how are they sealed?  What is your water source for spawning?

Your pics aren't particularly helpful, because the plate looks good, the jar looks good, the bag looks good I think, one tub looks good, but the other doesn't.  But they aren't even connected to each other.  We would need to see all the jars or bag that was used to spawn that green tub.  Also, was it a proven variety that you had success with before or something new?  Sometimes I carry from plate to tub a new variety and it goes horribly.  I think the plate is good, but I'm wrong...



Well I hope it is my grain prep because that could be easily addressed. And ya know in the beginning when I had way more success I soaked for 18 hours and got barely any burst grains. I got frustrated part way through this run and finally started posting this thread when it was too late to track a plate to a jar to a bag to a tub, but I will definitely be doing that on this thread for the next run with several different plates, and jar, etc.. from the same variety and from different varieties so please keep following along.

My water source for spawning is the tap water from a well that gets mixed into my coir and verm, it then gets pasteurized and loaded into a tub the next day. I do fill up a mister with tap water and lightly mist the walls of the tub and the top surface of the tub once it's spawned. Very lightly just to up the humidity in the tub right away. Then I leave it closed for several days until its fully colonized and open it up to check for contams and dry spots.

About these current pics, even though they aren't tracked from start to finish for a more proper diagnosis which I will do and post here next time... they are a perfect reflection of what has been going on for years now. Plates look fine, jars look fine, some tubs make it 1 or 2 flushes and half the others get thrown out before the first flush. Consistently every run regardless of the variety. I didn't cherry pick good looking plates or jars to post pics of. It's what everything always looks like..

Basically I don't have any proven varieties anymore. All the stuff that gave me success when I started growing is over 4 years old and has since been replaced with new stuff. When I started having problems and posted them here a year or so ago people told me my cultures were probably senescent and I should start new stuff. I've started several varieties from spores and have made transfers until the culture looks clean like in the initial pics on this thread and use those. I also have taken clones from multispore tubs and grow those out. Every single culture I have suffers the same plight, the same exact shade of green around the same time when fruiting. I haven't seen a unique contam in ages. When I started growing I'd see rhizopus, cobweb, different shades of green, etc.. Now it's always the same exact green on every different variety. Which has always made me suspect an overwhelming presence of one particular kind of pathogenic or parasitic mold in my home. But that could still come back to bad sterile technique or the old myth about spore loads in a home. I bombed my whole house before this run and it didn't seem to help though..


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


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OfflineSherlock Shrooms
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Re: Please help me get this right, contamination blues.. [Re: rickyswamps]
    #26853088 - 07/30/20 07:24 AM (3 years, 5 months ago)

Quote:

rickyswamps said:
You are overthinking your grain prep for sure.  They don't need to be soaked or rinsed, just boiled and hydrated.  A few burst grains are no big deal.  What kind of bags are you using (filter size) and how are they sealed?  What is your water source for spawning?

Your pics aren't particularly helpful, because the plate looks good, the jar looks good, the bag looks good I think, one tub looks good, but the other doesn't.  But they aren't even connected to each other.  We would need to see all the jars or bag that was used to spawn that green tub.  Also, was it a proven variety that you had success with before or something new?  Sometimes I carry from plate to tub a new variety and it goes horribly.  I think the plate is good, but I'm wrong...



The bags are unicorn bags with .2 micron filters and they are sealed with a 16 inch impulse sealer from FP, a nice one. Folks have mentioned tiny pinholes at the seal from excess heat but I've inspected every bag closely and I can't see any. They also hold their air volume well when mixing up. There's about 4 quarts per bag. I was loading them with 4 quarts then noc'ing them with 1 full quart but Pasty said to keep em at 4 quarts to avoid anaerobic bacteria, so I load them with 3.25 quarts and noc them with a full quart now.. I store them 2 bags per 64 quart cleaned out tub while colonizing to keep the spore load on the filters down.


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."


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Invisiblerickyswamps
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26853108 - 07/30/20 07:57 AM (3 years, 5 months ago)

A sub making  one or two flushes before contaming is normal.  Thats considered success.  Some people will say that they get 20 flushes with no contams, thats fantastic...but....IME, during the spring, summer, and fall, where I live, I get 2 flushes from very clean spawn.  Then a contam comes in.  This is normal.  Also, I get far less contams in shoeboxes because I think the sub stays a little drier. 

If you have a tub go bad before a pinset, then its to blame on your spawn and its most likely not grain prep.  Bacteria is easy to spot.  It gets wet and greasy looking.  Hidden fungal or white mold contams can be trickier. If a plate is growing slower than than others...toss it.  If it takes a lot longer for a jar or bag to colonize than your other ones...toss it.


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OfflineMycobro420
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Re: Please help me get this right, contamination blues.. [Re: Sherlock Shrooms]
    #26853111 - 07/30/20 08:01 AM (3 years, 5 months ago)

Have you ever took a agar plate open it for a minute infront of your flowhood.move it slowly all around the flowhood
Then parafilm it. Then leave it at room temp to make sure your flowhood is blowing steril air.


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OfflinePMBastian
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Re: Please help me get this right, contamination blues.. [Re: Mycobro420]
    #26853395 - 07/30/20 11:44 AM (3 years, 5 months ago)

Interesting thread, following.


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OfflineTweeq
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Re: Please help me get this right, contamination blues.. [Re: Mycobro420]
    #26853404 - 07/30/20 11:49 AM (3 years, 5 months ago)

I've been reading this whole thread and while Im also no expert:

Grainprep: Ive stopped rinsing, never used gypsum, soak for 12 hrs, boil untill first hull bursts

Sterilisation: the general concensus seems to be that 90 minutes @ 15 psi is the way to go. I think you are way oversterilizing those grains.

The problem seems to be your grains. Too many broken hulls and over sterilized they are too easy to get invaded by other stuff is my 2 cts


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Invisiblesandman420
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Re: Please help me get this right, contamination blues.. [Re: Tweeq]
    #26853459 - 07/30/20 12:31 PM (3 years, 5 months ago)

Spawn bags always have a lot of busted grains and are sterilized for 3-4 hours....Thats not it imo.


--------------------
- Sandbag Tek - How To Sterilize Spawn Bags - All About Static Pressure / Pressure Drop for DIY Flow Hoods - Sandman's LC Tek-

Marijuanaut escapes earth to cultivate - Grow-room is church temple of the new stoner breed


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OfflineTweeq
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Re: Please help me get this right, contamination blues.. [Re: sandman420]
    #26853642 - 07/30/20 01:51 PM (3 years, 5 months ago)

Quote:

sandman420 said:
Spawn bags always have a lot of busted grains and are sterilized for 3-4 hours....Thats not it imo.




Like I said, im no expert lol.

My process works for me at least.. sometimes with much worse looking agar than his.

Ill certainly hang around bc now Im curious af what it will turn out to be in the end


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OfflineMLPismyOPSEC
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Re: Please help me get this right, contamination blues.. [Re: Tweeq]
    #26853913 - 07/30/20 03:55 PM (3 years, 5 months ago)

Have you tried different brands of coir? What brand do you use? It has to be coming from something that has been constant in your technique, which makes me think either grain or coir. You say you bombed your house to clean it, what exactly did you do?


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