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Chronicles Of The Mean Green


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#1 macgyver

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Posted 09 October 2019 - 07:39 PM

To start this thread Ill provide some background. I never use grains only BRF/PF tek.
I started with B+ MS syringes to pf tek, with some success
I then used leftover MS to inoculate liquid cultures.
The liquid cultures appeared clean. (visible mycelium no discoloration, nothing floating on the surface)
Using these LC s I inoculated PF tek jars.
I crumbled 3 jars into a ziploc bag with sterile media.

After the rest of the jars are 100% as well as the bag, I spawned to a monotub.
I used Damnien's bucket tek and added 3TBS Gypsum, 1 TBS Olive oil, and 1 cup of spent coffee grounds (coffee grounds were freshly used and could very well be the reason for the Trich)
(got impatient and kept opening the bucket towards the end to check temp)
(Also an accidental low spawn to sub ratio)

Crumbled and mixed evenly and added a thin layer of sub over the top.

It looked good for 9 days
20190930_152107.jpg 20191002_220343.jpg


on the 10th day Trich appears (circled in picture) (Did not notice at first)
20191005_153852.jpg 20191005_153841.jpg

on the 11th day Trich takes over (picture taken outside, before DE-CON)
20191006_161852.jpg 20191006_161858.jpg

I quarantined this tub.

on the 13th day Trich sporulates (picture taken before outside, before DE-CON)
20191009_132011.jpg 20191009_132023.jpg 20191009_132020.jpg

In the same grow room I have another test ziploc bag, which I crumbled two stalled cakes and squirted a few cc's of LC over top. This was colonizing well(100%) in 5 days there was a bit of cobweb so I dunked it in a 10% peroxide solution for 3 hours then rolled in verm and spawned to a SGFC
20191005_150426.jpg 20191006_195105.jpg

I spawned the brick into a SGFC.
20191006_213734.jpg

2 days after spawning the brick im seeing this
20191009_155926.jpg

Could be bruising, guess I will know in 24 hours

Now in my mind there are 3 possibilities:
1. The bag was contaminated and got into the tub (not as likely).
2. the Trich from the tub got into the SGFC/bag.
3. My liquid cultures were bad making both end products bad.

My main questions are:
What is the likelihood that the LC was bad, but the cakes were flawless?
Can BRF cakes look wonderful for 2 full weeks and still harbor trich?
Or was it more likely, lazy pasteurization/coffee additives?

This has all caused me a lot of stress :blush: Lots of time and effort has gone into this and im looking for answers. Just want a little advice and peace of mind moving forward.
Thanks!


Edited by macgyver, 09 October 2019 - 07:59 PM.

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#2 FunG

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Posted 09 October 2019 - 11:05 PM

Liquid cultures are always questionable, you cant see mold spores in lc.

I think you can eliminate it as your primary culprit though since you did get two healthy cakes.

I'm going to say that it's the coffee grounds and oil additive, from my experience trying oils in bulk substrates they never worked out, not once, never.
Coffee requires a 1:1ratio to maximize success with it, the safer alternative that's just as nutritious as coffee grounds is worm casings.

Keep in mind, pfcakes can be crumbled at a 1:1 ratio with just plain coir and you'll get some really good results, it's when we tinker to raise nutritional value in bulk which isnt ment to be nutritional that trich makes its appearance....dont get me wrong, experimenting is great but if you just want fruit bodies then keep it simple and sweet.

You could try applying a salt paste to the area infected with trich to control the spread, your casing looks like it's about to pin....or at the very least, cutt off the infected area and plant it outside in a bed, you should get some fruits out of it still.

But yea man, if I were you I would just make diluted spore syringes by sucking up sterilized water after every 2cc's of use, you can stretch a dark syringe much further by doing so and avoid a potentially contaminated lc, they're good if you have those custom lids with the self healing injection port and filter but just making one, ones self usually results in disaster, at least it has for me.

Grain spawn masters are the way, you can see all growth and contaminates if any, the only problem is they only last 4generations before new ones have to be made...... not that at the fourth generation mark you probably wouldn't need anymore mushrooms lol

Hope I've been of some help

Take care
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#3 Billcoz

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Posted 10 October 2019 - 01:48 AM

I agree with FunG that trying to add nutrition isn't really worth it, and that LCs can be problematic for spotting contams, and from what I've read, coffee can be susceptible to contams.

 

I have used coffee a couple times without any contams, but it had no noticeable advantage over the plain substrate.

 

I have never used plain coir without at least some vermiculite, I do about a 50/50 verm/coir mix, maybe a little less verm, like 60/40 , and I sometimes sterilize it instead of doing the bucket tek and have noticed no difference in contam or growth rates.



#4 PJammer24

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Posted 10 October 2019 - 02:56 PM

if your cakes colonized completely and did not show signs of contamination prior to using them as spawn, your spawn is not the culprit.

 

I would suggest you do not use coffee grounds at all... They are notorious for inviting contamination.

 

How did you pasteurize your substrate... Improper pasteurization is the biggest cuplrit for causing contamination at that stage in my experience.

 

LC from multi-spore is a crap shoot. If you are making LC in a SAB or using a flow hood, they are not suspect at all... I have had maybe 1 bad LC in the last 50+. Agar to LC using a SAB is pretty damn reliable when done properly.

 

There are no ungerminated spores in your LC by the time you use it as one response suggested though there may be contaminate mycelium. Also, you can often see bacterial contamination on an LC... If it is not crystal clear (excluding the mycelium itself) then it is contaminated... A clean LC will allow you to see right through it... With time, you can develop the ability to spot contaminated LCs in many cases.

 

PF said that senescence never became an issue for him and he did much more than most people on this site. The deterioration of grain jar or anything else occurs with age and not the number of times you use a specific colony as a master. Someone please share some info on this if I am wrong but I don't think 4 "generations" out would be enough to cause a problem... I think we are looking at many many generations out before it becomes an issue but I have been wrong before.

 

Don't mess with trying to treat your trich with salt or H2o2 or anything else... If you can see green then there are MILLIONS of spores present and it is best to just get rid of it so they are not released into the air where you do your work.It sucks, but you are better off starting over or you may just cause yourself more issues down the road.


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#5 macgyver

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Posted 11 October 2019 - 12:33 PM

I removed the trich tub from my house, sprayed bleach on top of the sub to neutrilize the spores so I can minimize the chances of bringing any back in. Dumped the tub into a trash bag and double bagged it, sprayed the now empty tub with bleach again. Went back inside and immediately showered and changed.

 

The PF brick that is in my SGFC does not appear to have any trich, It was probably just bruising so I will up the misting and fae.

 

It is a relief to know that my spawn is clean, but made me think a little harder on keeping track of which LC i used to Noc what.

 

Going forward I will definitely skip the additives to the bulk substrate...

 

Also next time i will use a real pasteurization method instead of the bucket tek.

 

And a higher spawn to sub ratio.

 

In terms of the LCs i definitely want to learn more about AGAR and start using it.



#6 PJammer24

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Posted 11 October 2019 - 01:01 PM

I removed the trich tub from my house, sprayed bleach on top of the sub to neutrilize the spores so I can minimize the chances of bringing any back in. Dumped the tub into a trash bag and double bagged it, sprayed the now empty tub with bleach again. Went back inside and immediately showered and changed.

 

The PF brick that is in my SGFC does not appear to have any trich, It was probably just bruising so I will up the misting and fae.

 

It is a relief to know that my spawn is clean, but made me think a little harder on keeping track of which LC i used to Noc what.

 

Going forward I will definitely skip the additives to the bulk substrate...

 

Also next time i will use a real pasteurization method instead of the bucket tek.

 

And a higher spawn to sub ratio.

 

In terms of the LCs i definitely want to learn more about AGAR and start using it.

 

 

I would suggest using a different method but the bucket tek can give you good results when using substrates that are low nutrition like coir and verm. Its when you add highly nutritious additives like poo or in your case coffee grounds to the mix that the bucket tek will almost never be effective.



#7 macgyver

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Posted 11 October 2019 - 02:25 PM

 

I removed the trich tub from my house, sprayed bleach on top of the sub to neutrilize the spores so I can minimize the chances of bringing any back in. Dumped the tub into a trash bag and double bagged it, sprayed the now empty tub with bleach again. Went back inside and immediately showered and changed.

 

The PF brick that is in my SGFC does not appear to have any trich, It was probably just bruising so I will up the misting and fae.

 

It is a relief to know that my spawn is clean, but made me think a little harder on keeping track of which LC i used to Noc what.

 

Going forward I will definitely skip the additives to the bulk substrate...

 

Also next time i will use a real pasteurization method instead of the bucket tek.

 

And a higher spawn to sub ratio.

 

In terms of the LCs i definitely want to learn more about AGAR and start using it.

 

 

I would suggest using a different method but the bucket tek can give you good results when using substrates that are low nutrition like coir and verm. Its when you add highly nutritious additives like poo or in your case coffee grounds to the mix that the bucket tek will almost never be effective.

 

Right, Well on top of having the coffee grounds worm castings and olive oil, I kept opening the bucket every 5 minutes towards the end to take the temp because I was getting impatient and im sure that did not help either...



#8 FunG

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Posted 11 October 2019 - 05:06 PM

You used worm casings in the mix?

They're a lot more resistant to contamination then coffee grounds because of the beneficial microbes (whatever they're called)

Dung based substrates are not anymore susceptible to contamination then verm or coir, hpoo is the best bulk substrate one could use for cubensis cultivation because once again it too has those contaminate resistant microbes that fight off contams and ontop of that it can also have "firefang" fungus which is highly desired in aged horse manure since it prevents contamination even more so then the microbes and can be broken down by mycelium and used as a vital nutrient by the fungal colony. (Firefang is the only additive I'd recommend besides worm casings) and that would require a hpoo sub that either already has it running in it or is added by you through field gathering firefang nuggets.

I may sound crazy promoting something no one really talks about but yea, firefang is gold.

Keep in mind I normally use straight coir with grains and never have problems plus it yields very good but I do like to get fancy and tweak out a substrate from time to time but always keep it at a 1:1ratio no matter if its rye/wbs or popcorn.

You should see the flush's I'd get off straight coir using a 1:1ratio but with wbs, like 12oz's dry x that by however many monotubs....it was excessive growing.

I'm rambling at this point, damn stimulant drugs and I arnt chiming together so well on this post lol

Take care, and search firefang, its a interesting read up.

#9 Billcoz

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Posted 11 October 2019 - 11:01 PM

 

 

I removed the trich tub from my house, sprayed bleach on top of the sub to neutrilize the spores so I can minimize the chances of bringing any back in. Dumped the tub into a trash bag and double bagged it, sprayed the now empty tub with bleach again. Went back inside and immediately showered and changed.

 

The PF brick that is in my SGFC does not appear to have any trich, It was probably just bruising so I will up the misting and fae.

 

It is a relief to know that my spawn is clean, but made me think a little harder on keeping track of which LC i used to Noc what.

 

Going forward I will definitely skip the additives to the bulk substrate...

 

Also next time i will use a real pasteurization method instead of the bucket tek.

 

And a higher spawn to sub ratio.

 

In terms of the LCs i definitely want to learn more about AGAR and start using it.

 

 

I would suggest using a different method but the bucket tek can give you good results when using substrates that are low nutrition like coir and verm. Its when you add highly nutritious additives like poo or in your case coffee grounds to the mix that the bucket tek will almost never be effective.

 

Right, Well on top of having the coffee grounds worm castings and olive oil, I kept opening the bucket every 5 minutes towards the end to take the temp because I was getting impatient and im sure that did not help either...

 

Don't take the lid off of the bucket lol, the steam is what does the pasteurizing. When you dump the boiling hot water in the bucket over the sub mix/coir brick, let a little steam escape for 30 secs before you put the lid on so it doesn't blow off at your face, I've had it come off, but do get it on there as quick as possible and wrap it with a blanket or insulation, then don't open it until it's cool, or at least leave the lid on for a full 1-2 hours. That is plenty enough to pasteurize it and no real need to check temps, that's the appeal to the bucket tek in the first place. 


Edited by Billcoz, 11 October 2019 - 11:07 PM.


#10 macgyver

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Posted 12 October 2019 - 05:34 PM

Yeah I used a low amount of worm castings, 2 cups i believe and 3/4 cup of coffee.

 

But seems like its really not worth the hassle, for the coffee and i will probably will up the castings a bit.

 

And yeah the opening of the bucket plus fresh coffee grounds is 99.9% what made the mean green appear. I will probably up my spawn ratio as well just to try to colonize the mix quicker.

 

Thanks for the input guys!


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