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OfflineGrefa
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popcorn tek epic failure
    #21305155 - 02/20/15 05:36 PM (9 years, 2 months ago)

So I proceeded in soaking my popcorn and misread the tek which said to add white vinegar during the pressure cooking phase, not the soaking phase. Time to start over?

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Offlinespacechildo
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Re: popcorn tek epic failure [Re: Grefa]
    #21305187 - 02/20/15 05:46 PM (9 years, 2 months ago)

I have no idea but I found this quote which tells me you should try it anyway and report back!

Quote:

RogerRabbit said:
Quote:

2bafungi2 said:
It seems to be widely accepted that mycelium is most comfortable with a medium having a 5-6 pH value.





This is correct but mushroom metabolites will swing the pH acidic during colonization. If you're using well water with a high pH over 8 or so, a bit of vinegar or citric acid will lower it to neutral which is a good starting point.
RR




:shrug:

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OfflineGrefa
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Re: popcorn tek epic failure [Re: spacechildo]
    #21305196 - 02/20/15 05:49 PM (9 years, 2 months ago)

Thank you, I soaked in filtered water but the rinse will be well water. Thank you.


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Offliner00tuuu123
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Re: popcorn tek epic failure [Re: Grefa]
    #21305224 - 02/20/15 06:00 PM (9 years, 2 months ago)

How do you know the PH of your well water?


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InvisibleToadstool5
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Re: popcorn tek epic failure [Re: r00tuuu123]
    #21305817 - 02/20/15 08:11 PM (9 years, 2 months ago)

Quote:

How do you know the PH of your well water?




Test a small sample from the sink using litmus paper or an electronic pH meter.

Im not entirely sure its advisable to use a slightly acidic substrate. Fungi are not significantly affected by pH, whereas bacteria are. Bacteria hate slightly basic substrate (pH 8) but do great in slightly acidic substrate (pH 6.5-5.5) and mycellium metabolites lower pH.

In my opinion its better to start at pH 8-9 so bacteria is initially stalled, giving the culture time to establish itself. As the mycelium grows the pH will balance out to 6-5 (when you dont have to worry about bacteria). This protects you from contams while leaving a slightly acidic environment for the fully mature fungi to  take advantage of during fruiting.

Heres an article on pH and bacteria/fungi diversity:

Quote:

Soil bacterial and fungal communities across a pH gradient in an arable soil

Johannes Rousk1,7, Erland Bååth1, Philip C Brookes2, Christian L Lauber3, Catherine Lozupone4, J Gregory Caporaso4, Rob Knight4,5 and Noah Fierer3,6

1Department of Microbial Ecology, Lund University, Ecology Building, Lund, Sweden
2Soil Science Department, Rothamsted Research, Harpenden, Herts, UK
3Cooperative Institute for Research in Environmental Sciences, University of Colorado, Boulder, CO, USA
4Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO, USA
5Howard Hughes Medical Institute, University of Colorado, Boulder, CO, USA
6Department of Ecology and Evolutionary Biology, University of Colorado, Boulder, CO, USA
Correspondence: J Rousk, Department of Microbial Ecology, Lund University, Ecology Building, Sölvegatan 37, Lund 22362, Sweden. E-mail: j.rousk@bangor.ac.uk

7Current address: School of the Environment, Natural Resources and Geography, Bangor University, Bangor, Gwynedd, UK.

Received 17 December 2009; Revised 26 March 2010; Accepted 27 March 2010; Published online 6 May 2010.


Abstract

Soils collected across a long-term liming experiment (pH 4.0–8.3), in which variation in factors other than pH have been minimized, were used to investigate the direct influence of pH on the abundance and composition of the two major soil microbial taxa, fungi and bacteria. We hypothesized that bacterial communities would be more strongly influenced by pH than fungal communities. To determine the relative abundance of bacteria and fungi, we used quantitative PCR (qPCR), and to analyze the composition and diversity of the bacterial and fungal communities, we used a bar-coded pyrosequencing technique. Both the relative abundance and diversity of bacteria were positively related to pH, the latter nearly doubling between pH 4 and 8. In contrast, the relative abundance of fungi was unaffected by pH and fungal diversity was only weakly related with pH. The composition of the bacterial communities was closely defined by soil pH; there was as much variability in bacterial community composition across the 180-m distance of this liming experiment as across soils collected from a wide range of biomes in North and South America, emphasizing the dominance of pH in structuring bacterial communities. The apparent direct influence of pH on bacterial community composition is probably due to the narrow pH ranges for optimal growth of bacteria. Fungal community composition was less strongly affected by pH, which is consistent with pure culture studies, demonstrating that fungi generally exhibit wider pH ranges for optimal growth.




The entire article can be found at:
http://www.nature.com/ismej/journal/v4/n10/full/ismej201058a.html


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Invisibleblackdust
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Re: popcorn tek epic failure [Re: Toadstool5]
    #21306860 - 02/21/15 04:42 AM (9 years, 2 months ago)

:hehehe:





please do not change PH levels.

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OfflineGrefa
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Re: popcorn tek epic failure [Re: blackdust]
    #21332075 - 02/26/15 09:24 AM (9 years, 2 months ago)

its been almost a week since inoculation and not a single fuzz in any of my jars, thinking the vinegar ruined it. I doubt the local farmers market prices for oysters are going to change alot. Should I try and spawn another batch?


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InvisibleToadstool5
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Re: popcorn tek epic failure [Re: Grefa]
    #21332205 - 02/26/15 10:00 AM (9 years, 2 months ago)

I would wait another week just to be sure they aren't simply stalled. The acidity shouldnt have prevented the colonization so double check all your procedures.

If you have extra jars go for it! :thumbup:

Just out of curiosity, are you using spores, agar, or grain to inoculate the popcorn?


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Invisibleblackdust
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Re: popcorn tek epic failure [Re: Grefa]
    #21332276 - 02/26/15 10:13 AM (9 years, 2 months ago)

Quote:

bohdi said:
its been almost a week since inoculation and not a single fuzz in any of my jars, thinking the vinegar ruined it. I doubt the local farmers market prices for oysters are going to change alot. Should I try and spawn another batch?




I have a whole bunch of vinegar grains that were inoculated with slurry on 2/5/15. I see almost no growth. I see some white formations but it's slow. I can't say if it's mold or not. My temps have been slow so that contributes to the slowness but something should have happened by now.

get this.

  • 2 jars did not get sterilized
  • 2 jars were steamed for 90 min
  • 2 jars were PC for 90 min


and they all look the same. I dont smell bacteria but the vinegar may be covering that up. No slim either. Just grains that look the same over 2 weeks ago. I'm tempted to throw them out but I'm so damn curious to see what happens.

Here are some of the notes. They are not organized and I gave up on this project after a week. Now I am just waiting to see somthing then throw out. I really wanna know if they will get bacteria or mold because of the PH adjustment.

Project # 23

Normally, to generate grain spawn one would need a PC (pressure cooker) to get the grain sterilized enough for any clean inoculate to take affect and result in clean spawn. Clean spawn defined as achieving 1 dry oz of cubes per quart of spawn material. Liming grains have been reported to work but have yet to see physical proof. In my attempts, I will be working on getting a grain medium to produce 1 dry oz of cubes per quart of spawn as one would expect from using a PC.

- I have oven pasteurized wild bird seed flour and verm cakes along with coir/verm cakes with an inoculation of slurry. 1 PF cake was slurried into 9 small bread tins. All 9 tins fully colonized and proceeded to give a few flushes from all. Yield was bad but was expected per the conditions given. What did amaze me is I did this in a still air room and had all cakes fully colonized within days. I learned some good stuff from this project.

The only way I can see a way to get a whole grain medium to fully colonize is to use an LI for the inoculate which can be made from a PF cake that does not require a PC. 

IDEAS:
use of peroxide LOWER PH
Grain soak times
Adjusting PH of grains
Oven pasteurizing
Pickling lime RAISES PH
steam sterilization works but takes 8 hours…
Vinegar to lower PH
Bleach to raise PH


---
I believe the largest threat will be bacteria. If I can beat the bacteria then an LI should have no problem to colonize a spawn medium that is not as sterilized as a PC cycle will provide. 

THE REASON:
The results that whole grain spawn mediums provide seem to be a lot more favorable then use of grain flours. I wish to feel new learners of the hobby of the burden of having to buy a PC to get amazing results and to be able to use grains as a spawn medium. 

Conclusion:
I know their are a thousand way to skin a cat and many other people have found ways around this issue. In fact, Eat has shown that slurrying some PF cakes to a bulk sub with Hippy3 supercake has shown very favorable results. I just want to mess with this for my own entertainment and welcome any ideas or opinions. CHEERS!




Update: 2-4/15 Changing PF cake to a WBS jar

Need to hydrate WBS and drop ph to 4
PC some agar

• Sterilizing birdseed by baking or microwaving it will prevent sprouting, but some bird enthusiasts think that doing this destroys the seed's nutrients. If you want to try this method, spread the seed out in a single layer on a cookie tray, and then bake in the oven for eight to 10 minutes at 140 degrees Fahrenheit. Alternatively, put birdseed in a paper sack and microwave for five minutes.
• Put out seeds that birds in your area will eat. For example, striped sunflower seeds attract cardinals, woodpeckers, chickadees and doves. If you don't have those birds in your area, the seed is more likely to go to waste.

From <http://homeguides.sfgate.com/stop-birdseed-sprouting-70826.html>

Distilled white vinegar usually measures around pH 2.4, with a strength of 5%. Apple cider vinegar will have a pH of about 4.25 to 5.0.

From <https://www.google.com/webhp?sourceid=chrome-instant&ion=1&espv=2&ie=UTF-8#q=what%20is%20the%20ph%20of%20vinegar>



Reduction of Bacteria in Field Sponges after Chemical Treatment*


Treatment


Bleach- I :2.3 dilution Hydrogen  peroxide
lsopropyl alcohol Formula 409
I :2 dil ution undiluted
Ammonia Vinegar
*Unit of measure is CFU per sponge.
**Based on average count, determined by:

Gypsum is used to prevent PH swings
Vinegar will be used




Their is actually a thread at Topia where a guy used anti-bacterial soap on his grans and I think he got fruits. 


Hypothesis:
When we sterilize grains we are creating a window of opportunity for an inoculate to take root and beat competing organisms. That the speed of the inoculate determines the amount of sanitation a medium needs to flourish. Faster working inoculates like slurry or other LI/LC do not require an as aggressive sanitation method of a grain medium compared to inoculates of an agar wedge or MS.

Test 1:
Testing out a slurry of a PF cake to various grain jars of different sanitation methods. 

Parameters:
1. Work will be done in a still air room with all sterile methods used when working.
2. 1 PF cake will be put in a blender with 50ml peroxide and 150 ml of distilled water and blended by a couple short bursts.
3. The slurry will be used to inoculate 3 sets of jars with each set having 2 jars. A total of 6 jars will be made from the slurry.
4. Set 1 will have the WBS prep by hydrating for 8 hours then boiling. I will attempt to drop the ph of the grains to 4 or 5
5. Set 2 will have the WBS prep by hydrating for 8 hours then boiled then steamed for 90 min. I will attempt to drop the ph of the grains to 4 or 5
6. Set 3 will have the WBS preb by hydrating for 8 hours then boiled then pressure cooked for 90 min. I will attempt to drop the ph of the grains to 4 or 5

Conclusion:
I will be qualitatively judging the jars by color, growth, smell, and speed. By the end, I should have a better idea on what to focus on for the next preliminary project to reach the desired goal of this scientist. 




I may just be switching the PF cake with a colonized WBS jar I have thats been consolidating for about a week now just to speed up this test. If it works with the WBS grain jar then it should in theory be the same as a PF cake.



This starts in a few hours. Will be using a colonized WBS qt jar for the slurry of the jars. I will be using vinegar to lower the ph of the grain water for the grains to soak in. I attached a reading on a variety of disinfectants that were tested with sponges. Jars that pass the visual and smell test will be spawned to a oven pasteurized coir sub with a 1:1 ratio. Will also be adding in 50 ml of vinegar to 150Ml of distilled water for the slurry. More preliminary projects to be expected after and during this run. I will also be adding in gypsum to the soak water and grain jars to help stabilize the PH. 


Also, last run of unsterlized WBS has sprouts. I will be baking my WBS in the oven for 20 min to help prevent the WBS from sprouting on the unsterilzied WBS jars.



Nice. Set 3 of jars will be the ones non PC or steamed. All subs will be oven pasteurized. I just need to knock the bacteria back long enough for the 3 day colonization time. I have the WBS in vinegar now. Will have results soon. btw, got 20 lbs of WBS for 8 bucks!!!



I think they may all be going green. I can see growth starting and I'm not holding my breath. I also made a MS jar with the vinegar soak. MS jar may survive.

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OfflineGrefa
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Re: popcorn tek epic failure [Re: blackdust]
    #21335719 - 02/26/15 09:41 PM (9 years, 2 months ago)

Toad, I'm using liquid culture to inoculate the jars, from a supremely reliable source. Some aggressive mycelium if I get it it going. I'm going to do things a bit differently this next run of jars, perhaps I will have success after reading up more on jar prep. Going to put some vermiculite in the top of the jars this time


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OfflineGrefa
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Re: popcorn tek epic failure [Re: Grefa]
    #21340969 - 02/27/15 11:33 PM (9 years, 2 months ago)

http://imgur.com/VKuDMZp

Looks like a thowaway


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Invisibleblackdust
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Re: popcorn tek epic failure [Re: Grefa]
    #21342632 - 02/28/15 11:06 AM (9 years, 2 months ago)

Quote:

bohdi said:
http://imgur.com/VKuDMZp

Looks like a thowaway





1. How old is that jar?
2. How long was the soak?
3. What is the temperature is that jar is growing in?

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OfflineGrefa
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Re: popcorn tek epic failure [Re: blackdust]
    #21343236 - 02/28/15 01:08 PM (9 years, 2 months ago)

7 days old, soaked the grain for 24 hours, let it dry for an hour, PC'd and let cool overnight, inoculated in the most sterile conditions i could provide, lysol in air ducts taped off, mask, gloves, sleeves, glasses, put 2 cc at holes punched in 12,3,6 and 9 and put medical tape over the top, its been sitting for 7 days and its the only jar in the batch. Also this was my first run and as posted earlier white vinager got put in the soak so I thought i was abort anyways. Did another run of jars, this time i put a layer of verm in the top of the jars, when i dried the grain i spread it out on newspaper for an hour and a half, PC'd let it cool overnight, inoc'd with sterile conditions. Temperature for grain spawn is 72 degrees F


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Edited by Grefa (02/28/15 01:11 PM)

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Invisibleblackdust
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Re: popcorn tek epic failure [Re: Grefa]
    #21343299 - 02/28/15 01:23 PM (9 years, 2 months ago)

These jars are 23 days old. The 2 to the furthest right did not get PC'd or Steamed but a light boil. Also, The spray bottle is holding metabolites that are a few months old. I've been playing with it on my agar. :hehehe:

I still wonder how your jars contamed in 7 days and I  got some that did not get PC'd or steamed and look the same as 23 days ago when created. I'm so curisous to see what happens. Im'a knock em up again with some GLC in a week. I think the vinegar killed the spores but a LC could get around that.


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OfflineGrefa
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Re: popcorn tek epic failure [Re: blackdust]
    #21343544 - 02/28/15 02:36 PM (9 years, 2 months ago)

Good and bad news...its Trinch from moisture, which hopefully the myc will overrun


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OfflinetaGyo
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Re: popcorn tek epic failure [Re: Grefa]
    #21343652 - 02/28/15 03:02 PM (9 years, 2 months ago)

The myc does not overrun contams.

They co-exist and fight for nutrients in your bulk sub/grains. Those jars are fucked. A dry verm layer is for BRF, not grains. Dry verm does nothing in grains except soak up extra moisture.


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Re: popcorn tek epic failure [Re: taGyo]
    #21345219 - 02/28/15 05:36 PM (9 years, 2 months ago)

Yeah dont use a vermiculite layer for grains and the only time mycelium might "over-run" a contam is if it forms on the casing right before fruiting. RR has suggested using a sterile spoon and salt paste for any contams on casings but when your spawn is contaminated you are screwed because it will grow with the mycelium and continue to get worse.

You could try isolating some of the grains on agar but seeing as how you already have inoculate its not worth the hassle. :shrug:

Have you considered using a different grain? Some people have success with different grains. I've read white (soft) winter wheat has less endospores and rye berries are the traditional standard for mycology. Perhaps popcorn is not your grain?


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If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
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OfflineGrefa
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Re: popcorn tek epic failure [Re: Toadstool5]
    #21348125 - 03/01/15 12:50 PM (9 years, 2 months ago)

Popcorn was what was suggested from where I got the LC


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OfflineGrefa
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Re: popcorn tek epic failure [Re: Grefa]
    #21357143 - 03/03/15 04:17 PM (9 years, 2 months ago)

I think I figured out my problem, I was using refridgerated LC, and have no clue whether I just got the food for it or the myc. My best guess at this point is to take the LC outta the fridge let the myc form and reinoc.


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OfflineGreenRabbit
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Re: popcorn tek epic failure [Re: Toadstool5]
    #21357181 - 03/03/15 04:28 PM (9 years, 2 months ago)

Quote:

Toadstool5 said:
Quote:

How do you know the PH of your well water?




Bacteria hate slightly basic substrate (pH 8) but do great in slightly acidic substrate (pH 6.5-5.5) and mycellium metabolites lower pH.[/url]




Does this mean you could decrease the chances of contamination before colonization by using high pH water when doing a PF tek?

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