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An Errant Egret
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Registered: 11/12/09
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Egret's indoor Azurescens
#12237114 - 03/20/10 02:04 PM (14 years, 9 months ago) |
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This is something of a log, something of a tek, and the title is something of a misnomer - I'm currently trying to get my colonized substrate to fruit. As of March this whole process has taken me roughly 9 months from ordering spore syringes to the "almost there" stage. I've had an extremely low instance of (accidental) contamination while taking proper precautions, and while it took me a long time to do things right the process has gone EXTREMELY FAST when I've hit the sweet spots in every stage. If I can figure out the fruiting (which I think I have) I'm assuming the process can be broken down to about 2 months start to finish with virtually no risk of contamination.
In the Beginning...
I read the teks. I read the warnings not to modify them if you're a first timer. As I understood the fundamentals, the WHY of things, I felt that I needed only the magic number - the brown rice flour : water : vermiculite ratio - with which to forge my own path. My goal was to learn, not to copy, and while having mushrooms would be a plus having an understanding of mycology was the point. I picked azurescens because they're notoriously difficult, and because Spore Works sent me four syringes of them (and two cubensis syringes) for about $20. Yes, this is an AMAZING deal and is mostly due to them screwing up the order. It took me 2 months to get my syringes. At the same time, a friend sent me another 30 syringes from a successful harvest, so suddenly I'm swimming in raw materials.
After mixing my substrate and preparing my jars according to the MMGG, I decided to make my first change. To sterilize the jars/substrate I put them in the oven upside down for two hours at 300 degrees. This allowed me to do 144 jars in a single shot (I have a massive double oven and LOTS of jars - real men make their own salsa and know how to cook). The logic behind this change is that there would be no liquid loss since the rising water vapor would be trapped at the bottom of the inverted jar, and since contaminants are most likely to "fall" onto things keeping the holes of the jar facing downward as much as possible would further prevent contamination. Probably overkill, but hey, it worked. Two weeks later I had quarter-sized growth in ALL jars with absolutely no signs of contamination.
* NOTE: Cooking the jars like this melted the rubber seal onto the glass, which I didn't find out until later when the culture jars were ready to be birthed. Not a big deal really, but it's a bitch to detach the lids! Next time I'm facing the rubber away from the glass. Also, be sure to allow the jars to cool overnight since the center of the substrate will remain hot for quite some time - glass and vermiculite are excellent insulators! Furthermore, I'd suggest putting micropore tape over the holes in the lid just for that extra added protection against contaminants while still allowing gas exchange. My next grow will incorporate these and many more innovations that I'll share when I try them.
Snags, Breakthroughs, and Mistakes
From about two weeks to the 12 week point there was virtually no growth. The jars just sat there half colonized under my bed at about 70 degree temperatures. At this point I decided to get rid of all but 20 of them since I had an idea that required fewer jars and some friends who thought they could do better. My idea? An incubator. I had an old food dehydrator and put 8 of the jars into it, then covered that with a cardboard box and went to bed. About 6 hours later I went to check on my setup. The box was warm to the touch and the aquarium thermometer was maxed out at 120 degrees. I tore everything apart and went to bed, expecting to see a total loss the next morning. When I woke the growth had gone from a very vivid white to a light brown and had shrunk in size from about a half-dollar size to about a dime. I didn't give up, though - I was convinced my incubator idea was golden. In another two weeks I was ready to try again.
This time I picked up an in-line plug in thermostat, set it for 85 degrees, and tossed it in a cardboard box with an electric blanket and the culture jars. I placed the jars on top of some cardboard. Remember, kids, cardboard is contamination resistant and slightly breathable - you will want it underneath the holes in your jars at all times, assuming you store them "top down" or on top of your jars if you're into the "right side up" storage methodology. In another four weeks ALL jars went from vastly brown to entirely white - even those ravaged by the incubator version 1.0 which I'd written off as a complete loss. The cakes were even shrunk down about 20% in size as they tend to do when they're ready to be birthed.
* NOTE: I'm summarizing and incredibly hilarious experience for the sake of space - at this point I have 18 jars, not 20. After the dehydrator/incubator incident, I decided to rehydrate the jars since I had put them right-side-up and they'd dried out a bit. Two of the jars I submerged in water, but as I neglected to sterilize the water they became contaminated. The other 6 jars I rehydrated with a sterile syringe and sterile water, and they were fine.
I prepared for my next step by picking up disposable foil turkey pans with plastic lids. I also picked up some RED ALDERWOOD chips and began to soak these and my cardboard. A week later I was sterilizing the cardboard with the same two hours @ 300 degrees I used for the jars. The following day I sliced my cakes into thin slices to maximize surface contact between cake and cardboard, then put the pans into the incubator. Due to size restraints I could only do 4 pans. Everything went well and woodchips were sterilized/added in similar fashion once everything was fuzzy.
Pan Report
Pan A is my baby, and is the last "terrarium" I did. It's actually the dregs - the leftover woodships and the "slowest" culture jars, no cardboard. Surprisingly, this is the ONE pan I'm ready to try fruiting with. Chronologically, it's the last one of the four and wasn't created until pans B, C and D had woodchips added. But because it's the hero, it's pan A. Hooray for A!
Pans B and C are my guinea pig pans. After the cardboard was completely colonized and a light "sprinkling" of woodchips also became fully colonized I've intentionally and repeatedly contaminated them with various molds, bacteria, and other fungi over the last couple months. These pans will NOT be fruited, though I may sample the material to start a different colony in the future. I'm attempting to make an extremely contamination resistant strain. So far they've successfully defeated both Red and Green molds, E. Coli, orange slime mold, and are currently slowly winning a battle with a MASSIVE toxic black mold infestation. DO NOT ATTEMPT THIS. I have a dedicated clean room, thousands of dollars worth of professional lab equipment (so many schools/labs in these hard economic times), and plenty of experience working with common biological pathogens. If you don't even own a microscope or know what you're doing there isn't a point in RISKING YOUR LIFE trying this at home. That said, everything is progressing well.
Pan D is the fail pan, the first pan. It showed MASSIVE contamination within 48 hours and only went downhill from there. In all fairness, though, it's entirely my fault. I was in the basement when I prepared it and didn't sterilize ANYTHING except the cardboard - the pan, the knife, the cutting surface, the air... nothing was as clean as it should have been. Pan D went into the garbage the same week it was prepared. I lovingly refer to it as "my little abortion."
Don't Forget The Jars!
It's obvious to anyone growing that cardboard is pretty contamination resistant. In order to "save" the bits of white stuff stuck to the inside of the culture jars after birthing, I stuffed some soaked and sterilized cardboard into them, used tinfoil as a lid, and put those (upside down!) into the incubator. This seems to be an excellent way to save material - the cardboard became colonized and I surmise that I could realistically start another pan with them. I even added a few woodchips to some of the jars, and they too became colonized. This may be a great way to maintain "stored" cultures, as even after months of neglect they've remained viable.
Current Status
I'm ready to try fruiting pan A. I've got a dedicated fridge with a dedicated light, a wireless thermostat and a timer, and after running it for a week I've stabilized everything at 45 degrees with 6 hour on/off light cycles. I popped holes in the foil in pan A about 3/4 depth to allow air/CO2 exchange. After having it in the fridge for three days... NOTHING! I should at least have "dark zones" from what I understand. Thanks to these forums and TONS of research I've determined I may not be operating under ideal humidity conditions, that I may need to introduce unsterilized (casing) elements with nutrient/microbiological factors absent in sterilized substrate, and that I might want to consider slight temperature adjustments when the light changes (40 degrees light off, 45 on). There's still a lot to tweak and experiment with - I might even consider trying to fruit pans B or C just to see if their less sterile environment is key, though I wouldn't consider the fruit from those colonies to be safe for ingestion.
I tend to move slowly on my little mycology experiment, partially to analyze what adjustments do what exactly. I ordered my spores in August. Also, it's more of a hobby than a pursuit and I have much more exciting elements in my life that I devote the lion's share of my time to. By the end of this month I'll probably have a casing layer on. I'll keep this thread updated. The original "2 months start to finish" estimation is based on how much procrastination, experimentation, and error I can remove from the process. Hopefully this will help to nail down exactly WHY the indoor grows aren't working for people, since I can eventually isolate exactly what individual factors are working.
I would LOVE any questions or input.
-------------------- Ph. D. in particle physics, organic chemistry, molecular biology. Interdisciplinary minors in economics, psychology, infrastructure logistics. Over a dozen published works, two Nobel nominations, and accolades from twenty-seven world leaders.
So please, tell me in detail about the jack I don't know about shit that I might be privileged enough to glean a morsel of knowledge from your immense experience.
Edited by An Errant Egret (03/21/10 06:31 PM)
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TheAnimate
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Registered: 01/12/09
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Hmm... I haven't read the whole thing, but, 300 degrees is not really enough to sterilize a jar. You need a pressure cooker. You can get one that will do ten quart jars at once, this is still slower than your method, but it is the best way to do things. It may be that your jars didn't do anything because they weren't sterilized properly and the mycelia had to fight off contamination or bacteria. Don't turn your jars upside down, either. The water will get on the bottom and soak your filters, allowing contaminants to go right through the filter.
You should also look into liquid culture. Sounds like you used a lot of syringes, if you had made liquid culture, you could have used one syringe for all 120 jars. Liquid culture is faster as well, since it's already mycelia and doesn't have to germinate, besides the fact that you can use much more of it.
People used to use incubators but not anymore. I have seen a quote of RR himself calling incubators outdated by ten years or more.
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myceleus_rex
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Re: Egret's indoor Azurescens [Re: TheAnimate]
#12242258 - 03/21/10 01:30 PM (14 years, 9 months ago) |
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A PC only reaches 250F at 15 psi. Water boils at 212F at atmospheric pressure, so in an oven, even at 300F, the grains will not exceed 212F without drying them out, so yes, you need a PC. Not to be discouraging, but indoor azures are insanely ambitious for a "first timer".
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An Errant Egret
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My bad. I actually baked them for two hours, not one. I'll edit the first post to reflect this.
The internal temperature of the jar I'd jammed a thermometer into was at 300 degrees for 45 minutes, FAR more than is needed for adequate sterilization. I'm certain that the water in the jars boiled, but as I'd inverted the jars and the lids/holes were at the bottom the water vapor was trapped (water vapor rises).
I also neglected to mention that I allowed some of the jars to cool right-side-up to allow the water vapor to redistribute throughout the jars, but ultimately the results of doing this didn't show any particular benefit; while the growth in the right-side-up cooling jars was more uniform, it still became fully colonized in the same amount of time. I'm assuming the mycellium served some sort of capillary function to move water to drier areas of the substrate.
Please, feel free to be discouraging - I need a balancing opinion at this point since so far my results are EXTREMELY encouraging from what I understand. I'm fairly certain I've identified every single avenue of contamination I've experienced and can prevent my next generation from becoming contaminated entirely as well as cutting the "start to finish" time down to roughly 60 days. Based on experience, Azurescens can actually handle a lot more adversity than I've seen the mycology communities give them credit for. If I can induce fruiting (which I feel I've got the key to) I'll call this an overwhelming success.
Assuming success I'll make my own syringes, start from scratch, document everything, and make a proper tek with plenty of pictures. As for insanely ambitious, fungi are a ridiculously easy life form to understand with even a cursory high school education. Once you know WHY you're doing what the tek instructs you can fairly easily and safely modify things. I'll probably try a liquid culture at some point in the future, not because of a need to make my syringes go farther (I now have PLENTY of mycellium with which to spawn more cultures) but because it seems like an interesting alteration.
Growing mushrooms is like taking care of any living thing... understand its needs and why/how it needs them and keeping it alive/growing is a simple process. I don't think this is a particularly ambitious considering the massive amount of research I've done on the subject. I've had pets and plants which required living conditions much more difficult to create or support. Education in the scientific method (and to a lesser extent, the sciences in general) is the most vital thing any human being can learn. It teaches you a methodology to understand the world around you; an education in how to educate yourself, as it were.
TL;DR - don't respond to my posts if you haven't read the entire thing. You're both jumping to conclusions and telling me I'm doing it wrong when my methods are showing amazing results and a promising future.
-------------------- Ph. D. in particle physics, organic chemistry, molecular biology. Interdisciplinary minors in economics, psychology, infrastructure logistics. Over a dozen published works, two Nobel nominations, and accolades from twenty-seven world leaders.
So please, tell me in detail about the jack I don't know about shit that I might be privileged enough to glean a morsel of knowledge from your immense experience.
Edited by An Errant Egret (03/21/10 06:27 PM)
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myceleus_rex
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Registered: 07/01/09
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Well, all I can say is that I wish you the best of luck, and keep us posted 
You might like to know that there was a fellow a couple months back who had great success cultivating azures outdoors in Wisconsin. I don't have a link on hand, but I'm sure you can search it out.
Quote:
TL;DR - don't respond to my posts if you haven't read the entire thing. You're both jumping to conclusions and telling me I'm doing it wrong when my methods are showing amazing results and a promising future.
I did read your entire post, and I'm still skeptical about your sterilization procedure. In order to get the spawn jars to 300F without drying them out, you'd need to reach something around 30psi. in your jars (that is an educated guess, not a precise figure). Either you were very lucky they didn't rupture or the thermometer reading was inaccurate. I suspect the latter. I see no way that a container in which you "jammed a thermometer" could maintain the required pressure. I suspect that the heat outside with the dial housing was conducted down the thermometer to produce a false reading.
Edited by myceleus_rex (03/21/10 07:32 PM)
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DryGrain
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Sounds good. I'll be watching this thread. Good luck!
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libertaire
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Re: Egret's indoor Azurescens [Re: DryGrain]
#12245401 - 03/21/10 09:51 PM (14 years, 9 months ago) |
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I don't have time to read/respond to this in the proper fashion, but I will definitely be keeping an eye out for updates. You should post some pictures of your progress thus far.
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fig
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Re: Egret's indoor Azurescens [Re: libertaire]
#12245426 - 03/21/10 09:56 PM (14 years, 9 months ago) |
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_OttO_
Over Stimulated



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Re: Egret's indoor Azurescens [Re: TheAnimate]
#12245452 - 03/21/10 10:01 PM (14 years, 9 months ago) |
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That mycelium would fruit much better otudoors if your temps drop low enough there and if its the right season there.
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TheAnimate said: People used to use incubators but not anymore. I have seen a quote of RR himself calling incubators outdated by ten years or more.
An incubator sped up the colonisation time of my jars by more than double, when it was a cold time of year. In summer of course, there is little need for one.
I don't agree in the slightest that an incubator is outdated, depending of course on your climate and ambient room temp.
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libertaire
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Re: Egret's indoor Azurescens [Re: _OttO_]
#12247243 - 03/22/10 06:43 AM (14 years, 9 months ago) |
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Now that I have a bit more time to respond, I would recommend that you take that one successful tray that you have, sterilize/pasteurize 3 or 4 times that amount of wood chips, and use that tray to start an outdoor bed either in your backyard or a local park/woods. Even the most experienced and respected cultivators who have the most perfect climate controlled set ups have very little luck with indoor wood lover cultivation, so I fear it will be the same for you. For example, here's workman's attempt at it. Workman is the owner of the place you got your syringes, spore works:
http://www.shroomery.org/forums/showflat.php/Number/11680970#11680970
2 measly fruits, that's it. If you're willing to get only two fruits from the massive amount of effort and time, be my guest and continue trying to fruit it indoors, but you live in a climate where they will thrive, so why not give them the natural environment they need? Here's that thread of someone from your area that fruited them outdoors with great success:
http://www.shroomery.org/forums/showflat.php/Number/12208232#12208232
Secondly, the primary reason you lost the vast majority of your jars, and the remaining 20 took a shit load of time to colonize is because you used the oven at 300*F and a great deal of moisture was lost from your substrate, making it impossible for the mycelium to fully colonize the substrate. You could have easily used the oven with perfect success if you had only set it to 212*F, which as was mentioned is the temperature that moisture evaporates. This would have kept the correct moisture content and you would have had ass loads of spawn within a month. You live and learn though, and it seems like you're taking this all as a learning experience which is good.
I wish you luck in your project whatever you decide to do, definitely keep up updated with pictures.
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libertaire
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Re: Egret's indoor Azurescens [Re: libertaire]
#12247466 - 03/22/10 07:24 AM (14 years, 9 months ago) |
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Also, as I'm sure some will point out, sterilizing things in the oven is less effective because dry heat does not conduct itself nearly as well as wet heat (steam) does. For this purpose, perhaps on the next go around, not only should you only set the oven to 212*F, but perhaps also put the jars in trays that have water on the bottom to create wet heat, which will make it a lot easier to conduct the heat that's in the air.
Another option for future experiments is to try to use grains, which are much easier for building spawn then pf cakes because you can easily inoculate 10 new jars with one jar, thus making it possible to go from having 10 jars to 100 jars over night. One way you could do this with out a pressure cooker that people have had success with is doing an antibacterial soak over night and then steam sterilizing the jars. I suppose the oven sterilization method the way I suggested it would work here as well. Here's a link to the antibacterial sterilization:
http://forums.mycotopia.net/sterilization/21494-antibacterial-grain-soak-experiment.html
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An Errant Egret
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Re: Egret's indoor Azurescens [Re: libertaire]
#12247651 - 03/22/10 08:12 AM (14 years, 9 months ago) |
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Regarding pictures: I only took a few of the empty jars in the beginning. I had intended to log my experience from the get go, but after the two-to-twelve month mark where my culture jars were seemingly in suspended animation with only a small bit of initial growth I figured that it was going to be a failure and didn't bother. Lesson learned. I myself am generally a "pics or it didn't happen" type of guy, so I can understand any skepticism. After I'm done with this colony, I'll do another with full documentation. I'll also take pictures of my current setup IF I manage to get my colony to fruit.
Regarding Moisture: Why does it seem like nobody is understanding this? Water vapor rises, that's why we have clouds. As the bottom of the jar is solid and the jars were inverted, the rising water vapor was unable to escape the jars. Because of this, even WELL above boiling point, the moisture in the jars was unable to escape and my substrate ratio remained unchanged as a result. Water doesn't just magically escape an environment any way it can, it obeys the laws of physics like anything else. You can see this yourself by putting a bit of water in a pan, placing an inverted glass into the pan, and allowing the water to boil. Even at extreme temperatures long after the rest of the water boils away you'll have virtually the same amount remaining in vapor form (and condensate) in the glass. If you want to simulate the rest of the substrate materials in the jar, stick a sponge into the glass. After allowing it to cool you'll be able to extract that same amount of water out of the sponge. Don't use nonstick cookware, it damages the coating to be heated "dry".
Regarding Temperature: Any amateur cook will tell you that you don't just stick a thermometer into something and let it sit in the oven - that will indeed lead to a false temperature reading. I simply inserted the thermometer at 15 minute intervals to take my readings.
Regarding Indoor/Outdoor: When I started this project it was for learning purposes, and as a result I wanted a very controlled environment. Also, by the time I was ready to birth my culture jars there was snow on the ground here in Wisconsin (we have a wide range of temperature "zones" and I happen to be in one of the cooler ones). Additionally I have a dog and live in a neighborhood with a lot of kids, and not wanting to be the crotchety old man of the neighborhood I allow my yard to be considered fairly public property. It's pretty well traveled, and the last thing I need is a conspicuous patch of something I don't want trampled. Waiting for the proper outside conditions doesn't thrill me, either, though I may try an outdoor bed one of these days. Not to tell an experienced grower "you're doing it wrong" but personally I'd tweak Workman's formula a bit based on the dozens of other documented grows I've read about - ironically, I likely face the same situation. Maybe drop the temperature and alter the fir chips to a different hardwoord.
UPDATE I've got my casing layer. It's a mix of old maple and oak leaves I left on my lawn over the winter (it helps to better insulate the grass against the freezing temperatures - natural mulching) as well as some potting soil. I tossed in a very sparse sprinkling of grass seeds for shits and giggles. I've yet to adjust my light/temperature settings, but that's my next step should my current conditions not suffice.
-------------------- Ph. D. in particle physics, organic chemistry, molecular biology. Interdisciplinary minors in economics, psychology, infrastructure logistics. Over a dozen published works, two Nobel nominations, and accolades from twenty-seven world leaders.
So please, tell me in detail about the jack I don't know about shit that I might be privileged enough to glean a morsel of knowledge from your immense experience.
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libertaire
liberator



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Quote:
An Errant Egret said: Regarding pictures: I only took a few of the empty jars in the beginning. I had intended to log my experience from the get go, but after the two-to-twelve month mark where my culture jars were seemingly in suspended animation with only a small bit of initial growth I figured that it was going to be a failure and didn't bother. Lesson learned. I myself am generally a "pics or it didn't happen" type of guy, so I can understand any skepticism. After I'm done with this colony, I'll do another with full documentation. I'll also take pictures of my current setup IF I manage to get my colony to fruit.
Why not take pictures of what you have right now?
Quote:
Regarding Moisture: Why does it seem like nobody is understanding this? Water vapor rises, that's why we have clouds. As the bottom of the jar is solid and the jars were inverted, the rising water vapor was unable to escape the jars. Because of this, even WELL above boiling point, the moisture in the jars was unable to escape and my substrate ratio remained unchanged as a result. Water doesn't just magically escape an environment any way it can, it obeys the laws of physics like anything else. You can see this yourself by putting a bit of water in a pan, placing an inverted glass into the pan, and allowing the water to boil. Even at extreme temperatures long after the rest of the water boils away you'll have virtually the same amount remaining in vapor form (and condensate) in the glass. If you want to simulate the rest of the substrate materials in the jar, stick a sponge into the glass. After allowing it to cool you'll be able to extract that same amount of water out of the sponge. Don't use nonstick cookware, it damages the coating to be heated "dry".
The proof that your jars were dried out lies in the fact that none of them managed to fully colonize, which will only happen as a result of not enough moisture. Properly prepared, the pf tek formula should provide a jar with at least enough moisture to fully colonize, so either you didn't mix them right or your jars dried out.
I understand what you're saying about the moisture being trapped with the jars turned upside down, but I think you might be forgetting to take into account the fact that there are holes in the lids of your jars, and as the steam replaces the air and builds up in pressure, that pressurized steam will eventually try to find a way out of that pressurized environment, whether that out direction is above them or below them.
If you're really intent on using the oven to sterilize substrates, I've been pondering this question for a while, and have come up with what I think may be a good way to sterilize jars without that moisture being lost. I've never tried it out yet though, so if you're feeling adventurous, be my guest.
1.) Take two lids and drill a small hole in each one. 2.) Lay the lids with the rubber seal facing up on both of them, one lid on top of the other. You want to have the holes in the lids opposite of each other so that they will build the pressure, but still allow a small amount of air/steam to exit as needed. An optional addition I've thought of that might help is to put a layer of polyfil between the lids to allow the steam to escape more easily, but this may prove to be unnecessary or even undesirable in the long run. 3.) Fill the jars 1/2 way with pre-soaked grains and 3/4 of the way with water. 4.) Set the oven to 250*F (I believe the rubber seal melts at something like 275*F, so you should never go higher than that) with the jars in them, and once pre-heated, take them out.
The only thing is that this was designed with grains in mind, so that after the oven sterilization, you can take the top lid off, put a piece of polyfil in the bottom lid's hole as a filter, and then do a steam sterilization for an hour to an hour and half.
Quote:
Regarding Indoor/Outdoor: When I started this project it was for learning purposes, and as a result I wanted a very controlled environment. Also, by the time I was ready to birth my culture jars there was snow on the ground here in Wisconsin (we have a wide range of temperature "zones" and I happen to be in one of the cooler ones). Additionally I have a dog and live in a neighborhood with a lot of kids, and not wanting to be the crotchety old man of the neighborhood I allow my yard to be considered fairly public property. It's pretty well traveled, and the last thing I need is a conspicuous patch of something I don't want trampled. Waiting for the proper outside conditions doesn't thrill me, either, though I may try an outdoor bed one of these days. Not to tell an experienced grower "you're doing it wrong" but personally I'd tweak Workman's formula a bit based on the dozens of other documented grows I've read about - ironically, I likely face the same situation. Maybe drop the temperature and alter the fir chips to a different hardwoord.
That's understandable, I wish you luck.
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An Errant Egret
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Re: Egret's indoor Azurescens [Re: libertaire]
#12250457 - 03/22/10 06:14 PM (14 years, 9 months ago) |
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Quote:
libertaire said: Why not take pictures of what you have right now?
I had a meeting with my son's teachers tonight, had to cook dinner, do some yardwork, cook/clean up after dinner, rotate my car's tires, and do some other standard homeowner stuff. As mentioned before this is a hobby not a pursuit. I've set aside a chunk of my day tomorrow to photograph what I have so far, which is essentially foil turkey pans 3/4 filled with fuzzy woodchips. Nothing to "wow" about it. My guinea pig cultures (pans B and C) only show the slightest bit of the toxic black mold infestation anymore around the edges and it's really hard to see, so it might not show up too well in the pictures.
Quote:
The proof that your jars were dried out lies in the fact that none of them managed to fully colonize, which will only happen as a result of not enough moisture.
The jars colonized amazingly fast (from roughly four spots 1.5" diameter to fully ready to be birthed in a few weeks) once they were at the proper temperature in the incubator. The amount of moisture lost via my sterilization method can assumed to be negligible since - from what I've read - this is a fantastic rate of growth. It could also be that the the culture jars were exposed to too much light before they went into the incubator.
-------------------- Ph. D. in particle physics, organic chemistry, molecular biology. Interdisciplinary minors in economics, psychology, infrastructure logistics. Over a dozen published works, two Nobel nominations, and accolades from twenty-seven world leaders.
So please, tell me in detail about the jack I don't know about shit that I might be privileged enough to glean a morsel of knowledge from your immense experience.
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An Errant Egret
Writer


Registered: 11/12/09
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Here's some pics of what I've got at the moment, sorry for the terrible lighting conditions indoors.
pan A prior to the casing layer.

pan B showing what's left of the toxic black mold that it's fighting - yes, it looks green in the picture.

four unbirthed culture jars. The yellow is due to moisture buildup between the jar and the substrate - FAR from too dry for all you naysayers.

my casing layer mixture (a bit of potting soil, some maple/oak leaves from last autumn, and some water that I mixed in after the picture).

the micropore tape I used...

...and how it was applied to the air holes I punched in pan A (there are 12 holes total).

pan A assembled with the casing layer and a humidity sensor

a close up of my current humidity

pan A in its new home.
-------------------- Ph. D. in particle physics, organic chemistry, molecular biology. Interdisciplinary minors in economics, psychology, infrastructure logistics. Over a dozen published works, two Nobel nominations, and accolades from twenty-seven world leaders.
So please, tell me in detail about the jack I don't know about shit that I might be privileged enough to glean a morsel of knowledge from your immense experience.
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Flop Johnson
Praise Skatballah


Registered: 09/22/05
Posts: 13,789
Loc: TX
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Edited by Flop Johnson (07/31/11 10:41 PM)
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libertaire
liberator



Registered: 08/06/08
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Quote:
An Errant Egret said:Quote:
The proof that your jars were dried out lies in the fact that none of them managed to fully colonize, which will only happen as a result of not enough moisture.
The jars colonized amazingly fast (from roughly four spots 1.5" diameter to fully ready to be birthed in a few weeks) once they were at the proper temperature in the incubator. The amount of moisture lost via my sterilization method can assumed to be negligible since - from what I've read - this is a fantastic rate of growth. It could also be that the the culture jars were exposed to too much light before they went into the incubator.
Yes, but you said you rehydrated the 20 cakes you had left. If the oven hadn't dried them out, you wouldn't have had to do this and they would have reached full colonization on their own after 3 weeks or so. 70*F is indeed a bit low for incubation, but not grinding to a halt low, and they should have been fine at that temperature. I only colonize mine at 75*F.
All of that's irrelevant now though, and it looks really great overall, I hope you see some fruits very soon!
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jokefox
Top of the chain



Registered: 12/22/09
Posts: 6,231
Loc: never where I should be
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Re: Egret's indoor Azurescens [Re: libertaire]
#12260925 - 03/24/10 08:06 AM (14 years, 9 months ago) |
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fuck contams
nice to see you understand sterilizing and sterile procedures
i hope to fucking god this works for you good job mang
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An Errant Egret
Writer

Registered: 11/12/09
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Loc: Earth, for now.
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Re: Egret's indoor Azurescens [Re: libertaire]
#12261019 - 03/24/10 08:32 AM (14 years, 9 months ago) |
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Quote:
libertaire said: Yes, but you said you rehydrated the 20 cakes you had left.
Did I?
Quote:
An Errant Egret said: After the dehydrator/incubator incident, I decided to rehydrate the jars since I had put them right-side-up and they'd dried out a bit. Two of the jars I submerged in water, but as I neglected to sterilize the water they became contaminated. The other 6 jars I rehydrated with a sterile syringe and sterile water, and they were fine.
Emphasis added. I only rehydrated the 8 that went into the first incubator right-side-up and got dried out, and the other 12 actually colonized slightly faster than the rehydrated ones anyway.
I had an idea come to me in a dream last night. In one of the pans of my next generation I'm going to MIX the fully colonized woodchips with the casing layer and actually use far fewer chips, probably just enough to cover the bottom of the pan as opposed to a layer a few inches deep (something tells me the deeper stuff is essentially wasted). I'm also already soaking the woodchips so that they biodegrade slightly over the next months before I use them. The mixed substrate/casing material and degraded wood is meant to better mimic natural conditions, and along with the cycling humidity/temperature/light that I intend to implement based on a chart of conditions in Astoria I'm assuming the fruiting conditions will be better.
I also want to get a small chest freezer rather than an upright bar refrigerator. Cool air sinks, and every time I open the fridge door the cold air "pours" out. Plus I'll be able to try slight variations in the fruiting conditions between the 'frige and freezer to find what works better. But I'm getting ahead of myself - got to concentrate on the generation I have going right now. Probably in a month or two I'll start the culture jars for my next generation and make another indoor thread/tek. Maybe some of what I learn will even help with outdoor grows.
EDIT: I'm leaving town for a few days so there'll be no updates for a bit. According to my humidity sensor there's been no change over the last 36 hours so I'm assuming the lid of the pan is helping to retain enough moisture (there's water droplets on the inside, just like when it was incubating) that my colony won't entirely dehydrate while I'm gone - even if it drops to say 70% humidity I'll be fine with watering it when I get back.
Edited by An Errant Egret (03/24/10 08:50 AM)
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An Errant Egret
Writer

Registered: 11/12/09
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Last seen: 7 years, 2 months
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Yay Wisconsin! Since the refrigerator is in an uninsulated, unheated building it's somewhat at the mercy of outside temperatures. While I've been gone the temperature has been hovering in the teens to the mid 30's, quite a drastic change from the 50's the week prior. When I got home and checked the thermometer in the 'frige last night around 8pm it was at 33 degrees. As a result of the temperature being so low I doubt that there's been any progress over the last week - zero visible growth. Humidity remains around 90%. On the plus side I can put the tray out in the sun for the day now that I'm home, and the clear cover will probably have a sort of greenhouse effect.
I need something to do while I wait.
I was even considering migrating pan A to a completely outdoor bed, but that'd pretty much ruin the "indoor" focus of this project and since I'm out of town for half the week I wouldn't be able to control the growing environment as well as I should. I'm also thinking about using pan B or C to make an outdoor bed. B seems to have completely overcome the black mold infestation, but hasn't colonized the chips as fully as pan C which still has some traces of mold. I'm not so sure either culture would produce edible fruit considering how much potentially toxic material they've accumulated while eating up the various contaminants I've introduced. I'd always intended to simply sample B and C to begin fresh colonies in "clean" substrate, but I'm unsure that I'd be able to get enough mycellium growth in short enough time to catch the fruiting season before summer.
I got some more Azurescens spore syringes and am ready to start another generation from scratch to document start-to-finish progress - growing mycellium over the summer would mean that I wouldn't have to bother with the incubator. I'd make a "proper" step-by-step tek along the way (starting with the oven sterilization of culture jars), and probably make an outdoor bed out of that generation. The tek would then conclude with both indoor and outdoor fruiting methods, assuming both are successful. The only problem is that if I started now I'd be ready to begin the outdoor bed right around June by my estimate, so the tek would have MONTHS worth of idle time until I got to the fruiting season. As I'd like to document growth stage times based on ideal conditions and want to present the first version of the entire tek in finished form this route doesn't appeal to me. I also want to see if my current azures actually fruit before attempting again.
Another option is to try a different strain entirely while I'm waiting to see if the indoor Azures fruit successfully. I have one syringe each of 09/09 ps. cubensis (malabar), 10/09 ps. cubensis (treasure coast), and 07/08 ps. weilii. I'm not sure how long syringes remain viable in total darkness and 60-70 degree temperatures. Also, keep in mind that the Azures are my very first attempt at growing, and while I now know the basics of mycology I've spent most of my many MANY hours of research learning about Azurescens specifically. I'm mainly looking for the fastest culture-to-fruit strain as encouragement that I CAN grow mushies - from what I understand the cubes are pretty darned easy/fast, so I may try one of those. Anyone have any input?
-------------------- Ph. D. in particle physics, organic chemistry, molecular biology. Interdisciplinary minors in economics, psychology, infrastructure logistics. Over a dozen published works, two Nobel nominations, and accolades from twenty-seven world leaders.
So please, tell me in detail about the jack I don't know about shit that I might be privileged enough to glean a morsel of knowledge from your immense experience.
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