17 replies to this topic

[ShroomIfy59]

Ok the 10 pf cakes I nocked up more than 2.5 weeks ago are doing nothing the seem dead. Not sure what is going on. So I decided to make an LC so I could make sure the spores and syringe were able to produce mycelium

I followed the instructions from a beginner friendly LC post on shroomery... mostly. I don't have a PC but I do have a steamer setup that works pretty well. There is about 4 inches of water in the bottom and an upside down metal strainer holds up a round wire rack that's wrapped in foil to prevent splashing water. The tape below and above the water line allow me to check the temp using infrared thermometer. This setup keeps the temp in the steaming area of the pot at 99c. Not as good as a PC but seems to work pretty well. 

 

 

Here is what the distilled water and honey LC nocked up with 5cc of the JMF spore syringe that was 3 weeks old looks like after 4 days. 
I'm not sure I put enough honey in but there is definitely mycelium growing. 

 

 

I hope this helps jump start the next batch of PF cakes.   

Edited by ShroomIfy59, 12 September 2022 - 09:41 AM.

[ElPirana]

Good luck on the LC, looks good so far. Hopefully there’s no contamination hiding out in the jar. If it was me, I would transfer some mycelium out of there quickly to test it out, maybe make a few more jars of brf cakes.

[ShroomIfy59]

That's the plan. I'm going to give it a few days while I dump and clean some of my previous non performing cake jars. Then make 2 or 3 cakes and noc them up with the lc.

[MushLuvR]

If you swirl them a few times manually every day, the LC will be thick within 7-10 days.  I can tell you your LC is likely clean; I can see through the solution that doesn't have mycelium and that is always a good sign.  I find that within 2 days you will know, if it's cloudy, look closely and you will see particles floating, takes a sharp eye, but it's present and it's DIRTY.  

 

[ShroomIfy59]

I nocked up 2 fresh brf cakes yesterday from the LC. Hopefully this time I will see some growth. 

 

[ShroomIfy59]

Duplicate post

Edited by ShroomIfy59, 18 September 2022 - 07:28 PM.

[ShroomIfy59]

This is the growth after 3 days. It seems more grey than I remember from last year.

Does it look ok?

[attachment=è1198802:20220919_081527.jpg]

Attached Thumbnails

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Edited by ShroomIfy59, 18 September 2022 - 07:28 PM.

[Severian]

Difficult to tell but looks like mold to me

[MushLuvR]

Yeah tough at this stage to say for sure, but could be Wet Spot (slimy mold), probably most common right next to trich and Cobb Webb is last on that short list.  I have seen Pin mold on Agar from dirty syringes, but that's fairly rare for me to experience (being it isn't my initial work and bad source).  

Edited by MushLuvR, 19 September 2022 - 10:16 AM.

[ElPirana]

That doesn’t look good. I’d agree it’s likely mold.

Edited by ElPirana, 19 September 2022 - 11:16 AM.

[ShroomIfy59]

I will keep an eye on it for now. This batch of pf cakes has more water than before. I noticed that it was no where near field capacity when I mixed it so I added more water till it was close to field capacity. that combined with the LC injection maks it pretty moist in there. 

 

I will see what happens when more of the humidity inside the jar evaporates. When it shrinks back from the glass I should have a better idea what is going on. 

 

I took one to another room and slowly opened the lit and took a sniff. It smells like wet verm and brown rice. I don't smell anything odd. 

So I wait and see I guess.

 

Edit:

 

I think I found the problem. I used pond water from our fish tank for the first time in this brf tek. I boiled it for over an hour and filtered it through a coffee filter into a clean empty 1.5L pop bottle. It looked all clear and free of particulates at the time. I just checked the water left in the bottle and gave it a shake and now it's all cloudy with small chunks of grey stuff floating around. 

I guess I need to start over after sterilizing everything. 

Crap! 

Edited by ShroomIfy59, 19 September 2022 - 07:50 PM.

[Severian]

Pressure cookers here are really helpful

[ShroomIfy59]

I made 2 more pf cakes this time using distilled water to eliminate that as the problem. After 3 days still the same grey moldy mess. Yuk! 

 

I'm gonna get a few liter of chlorine bleach and distilled water and spray everything in my room I can.  Then scrap the LC I've been working from and start from scratch to see if I can get a clean LC.  to start from.

 

I was thinking I would just make the honey and distilled water solution and after pasteurization just put it on a brf paste and see if it grows the same mold. that way I can see if the honey is the source of contamination. I doubt that it is but I don't want to bet on it when I open a new spore print to start over.  

 

What a pain. Last year when I did it everything went perfectly for the first and second batch.  Now nothing seems to work as planned. 

 

At least I have other projects that I can work on that are going well.  

 

Pressure cookers here are really helpful

I ordered one today. Wife wants one too so It will do double duty for now and if it helps solve my problem I will get another PC for myself. 

 

Onward 
 

[ElPirana]

That pressure cooker will be a game changer for you, almost no point in these experiments if you can just wait for it to arrive. Your pf cakes and LC can easily be sterilized and you can narrow down any other possible source of contamination, if there is any.

[ShroomIfy59]

Ok the PC was delivered so I made 2 fresh brf cakes and a new LC jar to inoc with spores from a print I made last November. The high pressure setting on the PC got the temp up to about 119.5c according to my ir thermometer. 20 minutes under pressure at that temp then Turn off and cool down overnight.

This morning I setup my SAB and after cleaning it up and loading it with the new brf cakes, LC media, new sterile syringe, needle and spore print in a ziploc I sprayed lysol inside and closed up the SAB. Then sprayed the room with the lysol and put on my nitrile gloves sprayed alchohol on my hands before starting to work.

So I added spores to the LC jar using a flame sterilized exacto knife closed the LC jar tight.

Then I loaded the sterile syringe and needle with the Amazonian LC I made from spores weeks ago. This LC has been sealed was not exposed to the failed JMF LC from my previous attempt.



I nocked up the 2 brf cakes. Hopefully I will get mycelium this time not the nasty mold from the last attempt. Fingers crossed

[Severian]

Sooo if I'm reading you correctly, you used a spore print to inoculate LC?

If it wasn't a print made under sterile conditions, your LC is definitely going to contaminate because your spore print isn't sterile

As far as I am aware

The only way you can actually make spore print to LC work without contamination is if the mushroom was grown asepticly- as in allowed to fruit invitro and then opened in front of a flow hood.

The only other way to get a clean LC is to go spore-> agar -> LC.

[ShroomIfy59]

The Amazonian print was one I purchased last year and has been sealed since then. That's the LC in the last picture. It was innoc inside the SAB and was done first and closed up before I used a JMF spore syringe to innoc the contaminated JMF culture.  

The JMF print I used yesterday to try another JMF LC was a print I made from one of my first batches last year.

 

I fruit my cakes in a mini terrarium made from a 1.36kilo (3 pound) Skippy Jar. so each cake is isolated from the others while growing.

 

I made the print on foil sanitized with 70% ipa and placed under a sanitized plastic food container the foil and the box are on a clean and sanitized glass surface. I should have used my SAB I guess now that hind sight is 20/20.  So I guess I will hope for now and see what happens with both cultures.

 

If I'm gonna get into agar need to learn that process as well.

 

I will keep working on my environment and processes till I get it right. 

[ShroomIfy59]

The only way you can actually make spore print to LC work without contamination is if the mushroom was grown asepticly- as in allowed to fruit invitro and then opened in front of a flow hood.

The only other way to get a clean LC is to go spore-> agar -> LC.

 

 

I've been using the Chronic tec or a variation using Peanut butter jars. As I understand it not exactly in-vitro. 

I do appreciate your  feedback. Thanks for taking the time to help.