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#1 Saphroziac

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Posted 04 November 2020 - 03:06 PM

Hey guys,

 

I am depressed today.

 

1. Trichoderma is becoming a HUGE problem. I bought a flow food, and I buy all my substrate from a supplier for now, so I can more easily nail down where in my process it is going wrong. After sterilizing my jars of Rye for 1hr 30minutes to 3-4Hours at 15 PSI I let them cool overnight, and then inoculate from some of my LCs. I went big and lost out about 15 quart jars on a contaminated LC, and a month prior I lost 20 Jars to bacteria, and then tried the remaining 6 jars, on 3 bins of pasteurized coco coir/verm/cow poo, and they all had bacteria (you can tell when it isn't colonizing very well, and eventually contaminated out to trichoderma). It's been 4 months since I've had any mushrooms whatsoever. I prep the bins by washing them with dish soap, then spraying the entire thing with Hydrogen Peroxide H202, and then laying a plastic garbage bag down on the inside which I will also spray down with H202 then opening my bags of substrate in front of the Flow Hood, and pouring them into the bin. All with gloves on which I also sprayed wit H202. I then cover the top of the bin with Tin Foil to get a tighter seal around the inside, and poke holes in the top with a knife for FAE and cover with a few layers of Micropore Tape/Paper Tape. 

 

One bin contaminated out intensely all over after introducing O2 gas into the container to try and provide FAE without opening the bin, which makes me think the industrial oxygen isn't filtered before compression, and should instead be run through a filtering system then into the bin. The other 2 bins grew trichoderma, after a few days. This could be because either the jar was contaminated, or there wasn't a good enough seal with the tin foil, so perhaps I should just use the provided Bin Lid instead. I think I may skip the oxygen for now.

 

I am not intending to give up but after all of this I am starting to wonder if it is just not possible anymore to grow mushrooms in a house with 3 pets, a room with a carpet floor, and a garage with laundry going in an out constantly. I've pulled it off before, so...maybe more Mold Bombs?

 

2. The other Huge problem is Bacteria on Rye Berries/Hard Red Winter Wheat Berries especially few months ago. After months of waiting, watching, keeping faith the mycelium will continue to colonize and then it would slowly stop. I would open the jars and it would reek of wet shoe. Where it should instead smell of healthy mycelium. The source of this problem was that I was not drying my grains after soaking them for 18-24hrs. Causing a too high of a moisture content in the jars, and creating bacteria. Now after soaking I lay them out on a mesh screen suspended over a fan and dry them in the sun to try and evaporate and water on the surface of the grains. It could also be a contaminated liquid culture. That is how I am combating the bacteria problem. My only worry is that the grains are too dry; but they are inflated with water when I PC them, they're just dry on the outside.

 

I am also not sure my syringe filters work well as FAE filters because they clog with water after Pressure Cooking them and it can stay in there for weeks and you have to clear it with another syringe. Also the rubber self healing injection ports might be letting water in the through sides somehow. I've also trashed a few 3.5lb bags of Hard Red Berries because of a contaminated LC. I have 1 10lb bag of Millet that is working well and is constantly growing and is 50% colonized...so thats good I guess.

 

Here is the end of my struggles. Hopefully in a bout a month or so, these new LCs I've tested will work successfully. I tested them on 3 jars and there was no mold growing in them after 4-7 days. Hopefully Treasure Coast will work, and PF Classic. Albino Penis Envy Hybrid seems to be taking well it seems. My only worry is that my grains were too dry from being outside over a fan and in the sun for too long, and the mycelium doesn't get enough water to grow throughout the jar. I have 3 incubators filled with 16 inoculated quart jars each now. Maybe I should skip LC and just do straight Spore Syringe, but thats so expensive, and hard to do with a bulk grow. 

 

More patience, and more rigorous trial and error ahead.

Never give up! 


Edited by Saphroziac, 04 November 2020 - 03:14 PM.

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#2 FunG

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Posted 04 November 2020 - 03:41 PM

First off, stop using liquid culture (at least for the time being)

Liquid cultures are known to be problematic for people.

You need to test your spore syringe out before going to a liquid culture to ensure that it's not contaminated.

You can do so by inoculating some agar and waiting for growth to determine if the syringe is clean or not. Or if agar is unavailable just make up a spawn jar and test it on the jar by squirting the spore solution around the outer grains this way when germination occurs you will be able to see If there is any contamination. That's how I do it.

If the tester jar is successful, you can let it colonize and then grain to grain from it instead of using a liquid culture. G2g works better in my opinion, its safer, easier and just as quick as inoculating using a liquid culture.

Anyways, once you've eliminated the spore syringe as the culprit you could then proceed to try a lc again but I strongly advise you to just use grain to grain for the time being.

The carpet is a major problem, it houses tons of spores and bacteria that rise up in plooms whenever walked on, I had a apt with wall to wall carpeting and had the same experience as you with trich, healthy substrates would give into trich before the first flush....the apt was heavily contaminated with trich spores.

You can try a water/bleach mix in a spray bottle to mist you're work area that will cutt down on the mold levels significantly.

If you need a clean print of golden teachers I'll be happy to hook you up since I'll be cropping down some in the next week or so, and I know my prints are 100% clean....unless I take the lazy route and dont pressure cook the foil and containers but I wouldn't do that (ever again) lol

Also it's better to start from a trusted reliable source of spores rather then a no name vendor cause that's how these things happen.

Lesson learned, verify the spores are clean and free of mold/bacteria.

Also, it's ok if your rye is a little wet when loading the jars, as long as there is no pooling water in the bottom of the jar after loading then the excess will be absorbed by the rye grain after pressure cooking so you can stop worrying about drying them, I load wbs that's pretty wet all the time and like I said, as long as water doesnt pool the most you'll do is explode some grains.

Did I miss anything?
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#3 DickMoby

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Posted 04 November 2020 - 03:42 PM

Feels bad man.

 

The first thing I want to recommend is slow it down.

Figure the exact source of contamination out by doing very small scale tests, you fire a ton of resources and motivation by going all in all the time.

 

From what I've read it seems to be coming from the LCs.

While I think carpet floor is not optimal, especially with pets, I think it will still be possible to grow mushs in your home.

I've 6 cats and some of those little monsters are nasty, lovely bastards that puke behind things which you don't notice for long enough to grow mold on that pile of half digested goodness.

 

They shit more than they eat, I swear.. and with the air sometimes you could kill an elephant, still my agar is very clean even though everything is done in a SAB.

 

Maybe its the better option for now to switch to agar inoculations to see what you inoculate and to make sure the cultures are clean.

 

I personally sterilize all my bulk, mostly plain coir with from time to time some experimental additives or partly straw.

 

Some people say sterilizing bulk results in contaminations, not coir and straw.

I dont think your issues are coming from dirty bulks but maybe to eliminate that possibility that could be an option for you to try.

 

You mentioned millet.

300ml millet

175ml or gram water 

optional one spoon of spent coffe grounds

 

put into quart jars raw and sterilize - shake well after the jars are cooled.

no soak, no boil method.

 

Could safe you some time, it works good.

 

Can't help you with the lids, never used syringe filters.

 

Best of success for your future grows


Edited by DickMoby, 04 November 2020 - 04:41 PM.

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#4 TVCasualty

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Posted 04 November 2020 - 05:23 PM

I haven't used agar or LCs in years and almost never have a contamination problem (in a couple of different rooms in different dwellings). I pretty much stick with either BRF jars-->slurry--->grain--->bulk or BRF jars-->crumble to bulk. That also works for making and starting from isolates (blend a stem instead of a BRF cake).

 

Some people think BRF jars are obsolete or for really small grows but I think of them as sort of like 3-D petri dishes even though I can't really use them like colonized agar. They do a good job of revealing contamination (dirty spores), and if one jar is left uninoculated as a 'blank' then it can also reveal a problem with either the sterilization process or something in how the jars were handled afterward (that isn't related to contaminated spores or LC).

 

It's tricky when there's problems with both bacteria and mold/trich since molds and especially trich seems to be largely an airborne problem while bacteria seems to be largely a materials/sterilization/dirty spores problem. But that's speculative on my part (though based on experiencing both problems quite a bit and successfully addressing them as such).

 

I suspect that one of the reasons the slurry method works so consistently is the speed of colonization. It sure seems to be a distinct advantage, and I'm guessing is why Stamets titled his last book Mycelium Running. So starting a culture and watching it for a long time to see if anything unfortunate pops up might actually facilitate the unfortunate popping up, and it could be just about ready to show itself on agar or in a LC when it's decided it must be clean and used to inoculate a batch of grain. I've seen a very tiny tissue sample take 9 days to grow out enough on agar to be able to see with the naked eye. So maybe waiting and watching for 2 weeks or so would be another way to go since it's likely anything that would doom it would've revealed itself by then.

 

 

And grain used as spawn doesn't really need to be "fully" hydrated to work (or even close to it) since the myc spends so little time in the jar or bag that it has to be REALLY dry for inadequate moisture to become an issue (IMO). That's especially the case with bags, assuming they're filled with 3-4 qt. jars' worth of grain. Erring on the dry side has worked great for me.

 

Basically the more I treat them like land animals rather than semi-aquatic sauna dwellers the better they've responded. Bottled oxygen might provide enough oxygen but almost certainly not enough air flow, which is apparently essential to healthy growth, possibly by ensuring that enough oxygen actually reaches them at the rate they're consuming it. If you open the bin and breathe the air in it and feel you'd be fine with hanging out in that bin breathing that air then it's fine for them, too (that's how I check my bins if I don't detect any sign of contamination). If it seems stagnant and stifling then it is for the myc, too. I get the impression that they consume a lot more O2 than we tend to assume since the grows that work out the best for me have been the ones with the highest rate of FAE, or are outdoors.


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#5 Saphroziac

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Posted 04 November 2020 - 06:41 PM

Thanks for the extensive and articulate responses and advice everyone.  

 

I will begin from spore syringe again. I tested all my syringes out on separate rye jars today, as well as all my LCs. Hopefully in a few weeks we will see which ones are bad.

I also started a few Pan Cyan strains today. Hopefully these syringes are not contaminated. I also ordered a few Agar Plates to further test, I will proceed into agar, now that I have a flow hood. I think I will move the flow hood operation into the garage as well, where there are cement floors. I have a 4'x4' gorilla tent set up in there where I fruit and keep my incubators for now. 


Edited by Saphroziac, 04 November 2020 - 06:43 PM.

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#6 deepblueseawhale

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Posted 15 November 2020 - 01:10 PM

The only way I got beyond bacterial issues is when I did away with plastic garbage bags. For the life of me I could not get them clean and sterile enough.



#7 Saphroziac

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Posted 15 November 2020 - 07:06 PM

I tested all my Syringes and LCs on Agar Plates. Hopefully in a few weeks, I can see which are contaminated and which are not.

Treasure  Stack.jpg Treasure Coast.jpg PELC PANCAMBODI.jpg

 

I've also done a few Grain to Agar transfers with a strain that has taken to some Rye Berries.

Malabar G2A.jpg

 

I am sad because I think my Azurescens Woodlovers contaminated out. I even PC'd the woodchips for like 2-4 hours.

MOLD.jpg

 

Everything moves so sllloooowww. I tested some syringes like a week ago, and I can see the spores on the plate, but nothing is happening.


Edited by Saphroziac, 15 November 2020 - 07:08 PM.


#8 sandman

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Posted 15 November 2020 - 07:32 PM

Stop spraying everything with peroxide before you spawn. It's just making a wet place for bacteria to thrive when you do that because it's nearly all water and doesn't evaporate. Spray rubbing alcohol on your gloves occasionally. Alcohol is good for wiping down a bin too. You don't want to lay a trash bag liner in a wet tub. 

 

Ordering substrate, maybe they are sending you some shitty shit that goes to shit in the mail or not even prepared properly. It's already not ideal to spawn after several days old.

 

One grain per agar plate if that is what you are inoculating plates with, you don't want the grain all over the plate because there will be nowhere to grab a leading edge in a matter of days, just a big ol wad of mycelium. 


Edited by sandman, 15 November 2020 - 07:34 PM.

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#9 Saphroziac

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Posted 15 November 2020 - 07:53 PM

I am ordering from Santa Cruz Substrates. 

https://www.santacruzsubstrates.com/

 

Ok 1 Grain per Agar plate for now on. No more Hydrogen Peroxide.



#10 FunG

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Posted 15 November 2020 - 08:56 PM

Hydrogen peroxide is bad for mycelium, it weakens it and allows mold to get a foothold.

I use just plain old dawn dish soap for washing bins out, utensils get wrapped in foil and pressure cooked until their next use.

I know there is a hydrogen peroxide agar tek but it doesnt make sense to add peroxcide to agar before sterilizing since it will just revert back to h2o after pressure cooking.

#11 cujoloki

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Posted 16 November 2020 - 12:14 AM

I thinking is added in the agar cool down phase.

#12 fahtster

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Posted 16 November 2020 - 01:42 AM

If you have a flow hood, I’d ditch the lc for now as has been stated.. spore syringe to lc is a bad idea.. you should only use lc if you know for sure what you use to inoculate it is spotless. I’d say make pf cakes with the spore syringe and clone some fruits straight to pints of grain that you g2g to grain qts.

How are your tubs set up for FAE? If you’re using normal spawn ratios; 1:1, 1:2, 1:3, you shouldn’t have to cover your tubs with foil.. just put the normal lid on.. some FAE during colonization is fine.. the tub will colonize within 5 days and a little bit of light and FAE isn’t going to cause early pinning.. I implore you to start least try introducing fruiting conditions from the start on at least 1 tub time see how you like it.

Bacterial spawn can and does fruit but if you cut off the air while its colonizing, the bacteria is going to flourish and trich will take over.

I had a bunch of bacterial jars that looked like this
62A9E7C7-00BD-43FF-B92A-1D7A1803961B.jpeg

And because I put them into fruiting right away, they all put out flushes like this
06F6BFFD-8286-4017-9C78-8A6ECC17EE85.jpeg

And too wet grains don’t cause bacteria, bacteria causes bacteria.. I load burst grains and grains that have been sitting the strainer for only 20 minutes every time I run jars.. I purposefully cook my grains longer to get more burst grains because the myc loves the starch and rips through it.

Pf cakes —> clone fruit to grain—> 2g2 ( since you have a flowhood)—> tub with a normal lid and normal FAE (Ditch the foil and piped in o2.. fruit at birth.. doing that changed my life)

You don’t even have to grow the pf cakes out.. you can birth it early and just take piece of a leading edge of myc and put it to the grain pints for g2g

If you’re not using coir/verm, I highly suggest it.. it’s ridiculously easy and extremely contam resistant. Your house isn’t causing your tubs to contaminate, your contaminated grain is

Faht
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#13 Saphroziac

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Posted 16 November 2020 - 03:04 AM

I am thinking another critical mistake was the use of syringe filters. They only allow a hole for FAE that is about 3/16ths of an inch wide. Definitely not enough FAE, this is probably why I am having such slow colonization speeds.  Additionally, they clog up iwth water in the pressure cooker sometimes,, alowing for No FAE.

 

Do not use Syringe Filters as FAE Ports.


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#14 DrepsiLocybe

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Posted 16 November 2020 - 07:55 AM

I am thinking another critical mistake was the use of syringe filters. They only allow a hole for FAE that is about 3/16ths of an inch wide. Definitely not enough FAE, this is probably why I am having such slow colonization speeds. Additionally, they clog up iwth water in the pressure cooker sometimes,, alowing for No FAE.

Do not use Syringe Filters as FAE Ports.

this filters work way better than those syringe filters. ive been using them for almost a year now.

https://microppose.com/collections/all

#15 smellitstinknot

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Posted 16 November 2020 - 08:06 AM

Yes syringe filters are garbage for GE in my experience. Also when working with agar I'd recommend making at least 2-3 transfer from the original plate even if growth appears clean from the get go. Don't ever rush the agar work. It's always better to do a few extra transfers to ensure a clean culture than to toss a bunch of grain jars down the track. I go so far as to sniff my second to last transfer dish to ensure there's nothing hitching along on my mycelium or beneath. If the plate smells like mushrooms and growth looks clean I move the final dish to grains. Even so don't inoculate 20 jars at once. Just a few grain masters is plenty (I do 3) to test things out. If the mycelium races through the grains without irregular growth then go ahead and G2G to a larger number of jars. Back everything up so you don't have to start over from scratch if a project fails. Good luck :smile:


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#16 Saphroziac

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Posted 17 November 2020 - 03:38 PM

I have plenty of grain jars ready to go, just waiting on agar plates now.

125841398_406820077342472_3506675496499067391_n.jpg

 

The first agar plate! Pan Cambo Goliath! from an LC, I hope its not contaminated.

125534246_434535854612605_8135124155895123861_n.jpg



#17 Saphroziac

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Posted 18 November 2020 - 05:19 PM

First LC to get the axe!

125440957_1020617031750474_6246806837573953962_n.jpg


Edited by Saphroziac, 18 November 2020 - 05:19 PM.

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