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OfflineMrBoboFace
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Registered: 01/02/05
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Last seen: 10 years, 5 months
Experiment data - measured growth rate from Day 7 to Day 25
    #3572134 - 01/02/05 05:21 PM (19 years, 2 months ago)

Hey, everyone! I've done enough lurking and thought I'd toss my own experiences out. Be warned, it's a really long post.

I am a semi-first time grower using PF Tek (9 jars; duct-tape sealed-lid method) for an unknown spore print. I followed the technique fairly closely, using BRF, perlite, and distilled water for the substrate. The water was added slowly by hand until the entire substrate was damp (not wet). The jars were pressure-cooked (3 at a time) with a lid, band, and foil top for each jar.
While 9 of the 12 Ball jars were used for substrate, 3 jars were filled with distilled water and in one I placed an old, disassembled syringe (5 cc). This water was pressure-cooked and then used for the spore solution. I flushed the syringe three times into a waste cup, then expelled a fourth water sample into a glass salt shaker (microwave-boiling-water-sterilized), and used a flame-sterilized Swiss Army knife (cooled by sticking it in the syringe water) to scrape 1/4th a print into the glass salt shaker. My "clean room technique" was holding my breath a lot with the ventilation system off (no heat or A/C while making a spore syringe).

:frown: I should tell everyone that I tried to grow 12 jars of mycelium a few years ago using vermiculite, and the whole batch failed (bacterial contams. mostly) - I didn't use a pressure-cooker that time and did the "spray bleach water into the air in the bathroom" clean room technique. I did boil the jars for about an hour, but this either dried out the jars (all the steam escaped) or didn't heat them enough.

Back to the present...I have nearly reached the end of the incubation stage. The jars are all kept in their original Ball jar box, and they sit atop my always-on computer (a decent heat source). I kept a journal of the overall rate by which the mycelium was progressing, then did a graph of each day's growth rate.

Here is the graph:


Some important points (remember that action on Day X leads to reaction on Day X+1 or X+2):
1. Day 1 was inoculation day. Mycelium didn't spring up until Day 7.
2. On Day 7 the jars were moved from underneath my bed to the warm computer, and an insulating blanket was placed over them. You can see the general growth rate.
3. On Day 9 I was concerned that the substrate was getting too warm (31C) and I removed the blanket.
4. On Day 13 I put the blanket back on.
5. On Day 23 I removed the blanket again, for fear that the mycelium had been kept too warm (86F - I was overly cautious).

:thumbup: Warmth makes a big difference, guys! The best growth rates were when the jars were kept at 85F/31C. (this may be a "duh" note for the experienced growers, but it needs repeating for us newbies) Use the Tub-in-Tub method or do what I did and sit your box on a low-temperature heater. The Tub-in-Tub method is probably a better choice as it is more accurate and requires less maintenance.

:thumbup: Days of no growth is normal. The graph shows how a mycelium does a growth-halt-growth life cycle, with random peaks and dips for each jar. Patience will win out so long as you keep your babies warm.

:thumbup: Use a pressure cooker, even a cheap one. Just boiling the jars isn't enough. A pressure cooker prevents the substrate from drying out during boiling and gets the temperature high enough that it'll kill almost any bacteria or spores in the substrate. I'd say that 90-95% of contamination problems probably come not from a bad spore print but from not sterilizing the substrate.

:thumbup: Also, leave a layer of filler (vermic. or perlite) atop the jar before you put them in to pressure-cook. For this batch I had a 1" layer of perlite at the top, used a US$30 pressure cooker for 40-60 min, and so far have had zero contamination. Jars 3 and 8 were discarded, but only due to a total lack of mycelium growth for over a week and a half. Those jars were also free of contamination. I was likely too rash when I threw them out, though; they might've started growing again.

:alert: I'm not sure if the "remove tape and invert jar" method of improving air-exchange will work (with or without the pure perlite layer), but that will be tested in another batch.

:alert: In a third batch I will test temperature cycling - heating and cooling the mycelium within a 10-degree temperature range. Emulating the season change might encourage faster or healthier growth.

I do so love experimental biology.

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Offline13eetleJuice
the ghost with the most
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Registered: 10/28/04
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Re: Experiment data - measured growth rate from Day 7 to Day 25 [Re: MrBoboFace]
    #3572344 - 01/02/05 06:02 PM (19 years, 2 months ago)

Ok, I first read this and didn't want to touch this post with a 10' pole. I had hoped someone else would comment on it but it seems the general concensus is about the same. I'll start out by asking you a few questions and pointing out a few things that are likely to help you trememdously along the way.



Quote:

MrBoboFace said:
I am a semi-first time grower using PF Tek (9 jars; duct-tape sealed-lid method) for an unknown spore print.




Ok, either you are or you aren't a first time grower. How can you be a semi-first timer? There is only one first time. There's absolutely nothing wrong with it and you will be welcomed here no matter how much or little mycological experience you have under your belt.

You also mention that you are using an unknown sporeprint. I can only offer you my words of caution here. Be careful with things you do not understand or know the origin of.

Quote:

MrBoboFace said:
I followed the technique fairly closely, using BRF, perlite, and distilled water for the substrate.




Perlite is superheated volcanic glass and is used to regulate humidity in your fruiting chamber. It is not a suitable substrate. I assume you actually used vermiculite, but have gotten the terminology confused. Don't worry, as this is a common mistake of those new to mycology. If you did however use perlite and got results I can only say that I've never been introduced to this method. I'm sure it's possible but it's not really a decent substitute for vermiculite at all.




Quote:

MrBoboFace said:


Some important points (remember that action on Day X leads to reaction on Day X+1 or X+2):
1. Day 1 was inoculation day. Mycelium didn't spring up until Day 7.
2. On Day 7 the jars were moved from underneath my bed to the warm computer, and an insulating blanket was placed over them. You can see the general growth rate.
3. On Day 9 I was concerned that the substrate was getting too warm (31C) and I removed the blanket.
4. On Day 13 I put the blanket back on.
5. On Day 23 I removed the blanket again, for fear that the mycelium had been kept too warm (86F - I was overly cautious).




I suggest you make a separate graph for each jar and be sure that it is clearly labeled. I'm having a hard time making heads or tails of that graph.

Quote:

MrBoboFace said:
:thumbup: Days of no growth is normal. The graph shows how a mycelium does a growth-halt-growth life cycle, with random peaks and dips for each jar. Patience will win out so long as you keep your babies warm.




You will have MUCH less halts in the growth if the temperature is kept in a CONSTANT and OPTIMAL range of 84-86F. Mycelium hates changes in temperature (or anything for that matter). Consistency is the key.

Quote:

MrBoboFace said:
:thumbup: Use a pressure cooker, even a cheap one. Just boiling the jars isn't enough.




Boiling is actually quite sufficient when using BRF/Verm as a substrate. A PC is not needed if proper sanitary conditions are met throughout. When/if you move onto grains as a nutritious substrate you will need a PC or your battle with contams with be a long near futile one.

That's all I have time for right now. I'm sure others will have more to say in subsequent posts. I don't want to seem as if I'm condemning experimentation. I support it, fully. I only ask that you save yourself some time and read up on some of the teks and faq's available here at the shroomery. Many of the questions you seek to answer through experimentation have already been answered multiple times by those with decades of experience in mycology. Good luck to you and may this be a fruitful hobby for you. :smile:


--------------------

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OfflineMrBoboFace
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Registered: 01/02/05
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Re: Experiment data - measured growth rate from Day 7 to Day 25 [Re: 13eetleJuice]
    #3572675 - 01/02/05 07:03 PM (19 years, 2 months ago)

I'll do my best to respond to everything you mentioned.

Quote:

13eetleJuice said:
Ok, either you are or you aren't a first time grower. How can you be a semi-first timer? There is only one first time. There's absolutely nothing wrong with it and you will be welcomed here no matter how much or little mycological experience you have under your belt.



I used the term "semi-first time grower" as I had attempted to grow once before but got zero mycellium growth and tons of contamination. This is simply the first time I've actually grown a mycellium patch; that's all I was trying to convey.

Quote:

13eetleJuice said:
You also mention that you are using an unknown sporeprint. I can only offer you my words of caution here. Be careful with things you do not understand or know the origin of.



The spore print is unknown and therefore it is not to be grown for consumption. This is an experiment in growing mycellium; the cakes are to be discarded once full colonization is done. A waste, perhaps, but "when in doubt, throw it out".

Quote:

13eetleJuice said:
Quote:

MrBoboFace said:
I followed the technique fairly closely, using BRF, perlite, and distilled water for the substrate.




Perlite is superheated volcanic glass and is used to regulate humidity in your fruiting chamber. It is not a suitable substrate. I assume you actually used vermiculite, but have gotten the terminology confused. Don't worry, as this is a common mistake of those new to mycology. If you did however use perlite and got results I can only say that I've never been introduced to this method. I'm sure it's possible but it's not really a decent substitute for vermiculite at all.




I am not confused in the terminology. I did indeed use perlite for my substrate, not vermiculite (chipped mica). The perlite has done well so far. I know that perlite is used primarily to regulate humidity within a fruiting chamber, but I purchased the perlite as a substitute for the vermiculite that was needed to act as filler for the substrate. There has been substantial condensation on some of the jar walls, however, which may be a side effect of using perlite instead of the water-absorbing vermiculite. Vermiculite may encourage faster growth, however. I will need to test this.

Quote:

13eetleJuice said:
I suggest you make a separate graph for each jar and be sure that it is clearly labeled. I'm having a hard time making heads or tails of that graph.



And here we go:

1. Day 1 was inoculation day. Mycelium didn't appear until Day 7.
2. On Day 7 the jars were moved from underneath my bed to the warm computer, and an insulating blanket was placed over them. You can see the general growth rate.
3. On Day 9 I was concerned that the substrate was getting too warm (31C) and I removed the blanket.
4. On Day 13 I put the blanket back on.
5. On Day 23 I removed the blanket again, for fear that the mycelium had been kept too warm (86F - I was overly cautious).

Quote:

13eetleJuice said:
You will have MUCH less halts in the growth if the temperature is kept in a CONSTANT and OPTIMAL range of 84-86F. Mycelium hates changes in temperature (or anything for that matter). Consistency is the key.



I see. So you are saying that halted growth is not normal, then. Perhaps my next experiment should involve two batches: one with fluctuating temperatures, and another in a Tub-in-Tub setup with a constant temperature. Thanks for the tip!

Quote:

13eetleJuice said:
Quote:

MrBoboFace said:
:thumbup: Use a pressure cooker, even a cheap one. Just boiling the jars isn't enough.



Boiling is actually quite sufficient when using BRF/Verm as a substrate. A PC is not needed if proper sanitary conditions are met throughout.



Not in my past experience. Many newbie growers, such as myself, have not developed the necessary methods to maintain sanitary conditions. I make the suggestion of using a pressure cooker as it will assist the new grower in avoiding contamination.
My first time using a pressure cooker resulted in zero contamination, while a previous attempt without a PC resulted in total bacterial contamination. I maintain my statement that a pressure cooker is very helpful in reducing or eliminating contamination potential, even with the relatively clean BRF/vermiculite mix.

Quote:

13eetleJuice said:
Many of the questions you seek to answer through experimentation have already been answered multiple times by those with decades of experience in mycology. Good luck to you and may this be a fruitful hobby for you. :smile:




Oh, I realize that most of what I'm mentioning here is already covered in the FAQ and that my "experimental methods" are likewise fully tested and proven/disproven within the FAQ, by people with far more experience than me. However, all of these suggestions/warnings/methods should be able to be replicated by another grower. I'm testing the theorem. If I can prove a known fact, then I feel more confident in proving/disproving an unknown method.

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Offline13eetleJuice
the ghost with the most
Male

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Registered: 10/28/04
Posts: 2,253
Loc: 6' under pushin up shroom...
Last seen: 5 years, 4 months
Re: Experiment data - measured growth rate from Day 7 to Day 25 [Re: MrBoboFace]
    #3572823 - 01/02/05 07:48 PM (19 years, 2 months ago)

Looks like you're off to a good start then. I see what you mean by semi-first timer now. :thumbup: I didn't quite read far enough into what you said the first time. Oh, and thanks for thinning the graph out a bit for me. I was having a hard time distinguishing one line from the next. Just out of curiosity, what unit of measure did you use for growth rate? Did you just observe it daily and quantify it on a scale of one to ten?

Be aware that it will take longer for the myc to fully colonize if you look at your jars every day. This is something you can test as well, if you are inclined to do so. Try letting a control group of jars sit undisturbed for 15 days while checking the other group for growth and noting it's progress each day. On day 15 check the control group for the first time and see how much further along they are in comparison.


--------------------

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OfflineMrBoboFace
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Registered: 01/02/05
Posts: 5
Loc: North America
Last seen: 10 years, 5 months
Re: Experiment data - measured growth rate from Day 7 to Day 25 [Re: 13eetleJuice]
    #3572995 - 01/02/05 08:18 PM (19 years, 2 months ago)

For the scale of measurement, I looked at each side of the jar (4 sides total), and recorded a rough estimate in % of each side's growth. I averaged all 4 sides for the graph. I've since realized that such a method is inaccurate, so I'll find another method of measurement that involves "measurements", not "eyeballing".
That's a good idea, BTW. That would probably show me that a daily measurement would interfere with the experiment. Thanks again!

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