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owntharabbithole
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Need some advice for timing transfers and spotting contams?
#26546820 - 03/20/20 02:22 PM (3 years, 10 months ago) |
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I've been wanting to start growing again after a few PF Teks in my early 20's. The gist is that I wanted to try stepping up a bit and like many others, love tech & automation and have seen the arguments both for and against automation. Long story short, I want to dabble and work towards a proof of concept. So far I obtained spores from didntreadrules.com, was supposed to get 1 PE syringe and 1 pan cyan Jamaican print, however, they were out of that print so without asking to specify they just sent me the syringe of the cyans too. I've been reading and watching everything I could on the steps to the next level grow and was able to build a pretty badass (IMO) portable flow hood that uses an inline duct fan to get the CFM to 100 (actual CFM 107). I created 4 32qt tubs for fruiting and modified a Crane Drop humidifier to handle all 4 tubs with the use of "t's", although I don't expect it to run all the time.
 To start my flow hood was set up in the doorway of a closet to minimize the airflow around the environment. After spraying and wiping it all down, I used the syringes with pre-poured PDA agar dishes I sourced from Etsy. Next time I'd like to pour my own but I wanted to eliminate at least 1 contam vector and cut my startup work a tiny bit with the plan to make half Penis Envy and half Pan Cyan Jamaican. As for my noob agar procedure lol, I did just one ms drop off of an inoculating loop I made from 18ga SS vape wire that I twisted with a drill and secured in an Exacto knife handle and sterilized. I've never done agar and I def think I need practice so I don't gouge it lol. I don't know if the loop was too big, wire too thin, agar too soft, but when I tried to streak the various lines and patterns it would catch and dig in to the agar. Nonetheless, I flamed the tip in between each streak.
 Currently, for incubation I have all the dishes inside a smaller tub on top of a dry towel, then placed inside one of the 32qt tubs on top of a seedling mat that's controlled by an Inkbird temp/humid controller keeping it within 5 degrees of 83. Currently, there aren't plans for the Inkbird hygrometer; I imagine I could hook a fan up to it but again, I doubt on the small scale it'd be needed much anyway. They have been incubating for 11 days and now im HAPPY to say, I have lots of growth, both good and bad, and likely need to transfer but need to make sure im not grabbing anything im not supposed to lol. The green colonies are obvious but want to be sure the others are mycelium before doing all the work. I thought the pan cyans weren't working but then I spotted the thinner and almost transparent look of it as it starts. Some plates still have absolutely nothing still. Any suggestions or feedback regarding the transfers are greatly appreciated. I've read to make sure they are properly timed, before they hit the edge of the plate, and to look for the most characteristic sections to transfer to the new dishes. I think its time for some and the PE's look decent too, don't they? Im excited and grateful to be a part of this community ;-p
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26546842 - 03/20/20 02:29 PM (3 years, 10 months ago) |
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So I checked again today and decided to pull the dishes that still have no growth. Some of these look really nice, but admittedly im still a noob. From what little I know, the single clean circular dishes are great to keep, the mold one looks to have good growth sector to use, but what about the streak dishes with multiple spots? I think there is clean growth to get out of it but should I bother since I have the other better dishes? On the image of 5 plates, most look thin with the exception of the lower 2 and not in the wispy "good" way. Does my assesment sound somewhat accurate? Also, on the same 5 plate image the lower right PE dish seems to be yellowing slightly. I know that can be a lot of things, so my question is how do you rule out Aspergillus etc if it all can look just like mycelium lol?
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26546852 - 03/20/20 02:34 PM (3 years, 10 months ago) |
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Wonderful your a mad scientist you remind me of myself lol I did all sorts of stuff like this back in the day. The flowhood build is really unique! Keep puttering through it but you may wanna just check out how mono tubs are done now. It's a very set it and forget it thing that works great đ
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26546876 - 03/20/20 02:40 PM (3 years, 10 months ago) |
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Quote:
owntharabbithole said: So I checked again today and decided to pull the dishes that still have no growth. Some of these look really nice, but admittedly im still a noob. From what little I know, the single clean circular dishes are great to keep, the mold one looks to have good growth sector to use, but what about the streak dishes with multiple spots? I think there is clean growth to get out of it but should I bother since I have the other better dishes? On the image of 5 plates, most look thin with the exception of the lower 2 and not in the wispy "good" way. Does my assesment sound somewhat accurate? Also, on the same 5 plate image the lower right PE dish seems to be yellowing slightly. I know that can be a lot of things, so my question is how do you rule out Aspergillus etc if it all can look just like mycelium lol?

Those PE dishes that are non streaked and just round growth look good. Transfer the ropy rhizo leading edges to new plates. I dunno if I'd mess with the streaked ones much but you could try a transfer from them. You will see that mycelium just has a different growth rate and look than the white contams growths.
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26547746 - 03/20/20 10:32 PM (3 years, 10 months ago) |
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Quote:
rumfor69 said:
Quote:
owntharabbithole said: So I checked again today and decided to pull the dishes that still have no growth. Some of these look really nice, but admittedly im still a noob. From what little I know, the single clean circular dishes are great to keep, the mold one looks to have good growth sector to use, but what about the streak dishes with multiple spots? I think there is clean growth to get out of it but should I bother since I have the other better dishes? On the image of 5 plates, most look thin with the exception of the lower 2 and not in the wispy "good" way. Does my assesment sound somewhat accurate? Also, on the same 5 plate image the lower right PE dish seems to be yellowing slightly. I know that can be a lot of things, so my question is how do you rule out Aspergillus etc if it all can look just like mycelium lol?

Those PE dishes that are non streaked and just round growth look good. Transfer the ropy rhizo leading edges to new plates. I dunno if I'd mess with the streaked ones much but you could try a transfer from them. You will see that mycelium just has a different growth rate and look than the white contams growths.
Thanks, good to know I'll key my eye out for that. I did the mist and fan back in the day and the new monotubs look sweet. This is serving as a playground for testing different setups right now starting with a tight, closet space in an apt. Eventually making its way to a custom outfitted basement running something like a Rasberry Pi and https://kylegabriel.com/projects/2015/02/automated-mushroom-cultivation.html https://github.com/kizniche/Mycodo
Looking at this as reference:  You were suggesting I transfer from the outer ring of #3 which is a Penis Envy sample, and I imagine #2 would be sufficient for the pan cyans sample. As for the rest, what would you guys do? #1 has nice growth but seems very light. #5 obviously streaked out, so they all look good but nothing to write home about.....trash it? Same with #6, it's nice but looks obviously slower growth in most regards.
So I pick one or two dishes and take the samples I want, and then what? Throw them away?
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26547881 - 03/21/20 12:28 AM (3 years, 10 months ago) |
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I would do transfers from all of them that you can just to see what they do. #1 could end up becoming thicker or who knows what else. And you can't really have too much myce anyhow plus never know if one goes bad so it's good to have copies.
They can be stored in the fridge for a long time so don't just throw them out immediately. Wait until new plates grow clean before getting rid of old ones
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26577781 - 04/04/20 05:39 PM (3 years, 9 months ago) |
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I had started some new plates and did some transfers, but... Can a mod merge my two posts? https://www.shroomery.org/forums/showflat.php/Number/26577643
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26577926 - 04/04/20 07:18 PM (3 years, 9 months ago) |
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Second row down, second plate in, that 4-5 o'clock position. You need to transfer just a little piece of that ropy thick rhizo branch to another dish. That is definitely one to isolate and what you're looking for!
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26577932 - 04/04/20 07:22 PM (3 years, 9 months ago) |
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I'd make two transfers from right here to two separate plates
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26593866 - 04/11/20 05:07 PM (3 years, 9 months ago) |
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Back with updates, hope everyone is good during amidst all this craziness:) I took your advice on the transfers and while I had everything set up I tried some more pan spores to agar via swab since I don't seem to be getting the fast wispy growth. I've lowered the temps to max 80. I knew I wanted to get to play with LC so with the extra time I took the House of Hydro fan I never used and made a magnetic stirrer. For the rye inoculations, I used sterile syringes to make myc water for half of them, the other half were with agar wedges. I wanted to see the difference for myself:) They have been going for 11 days and seem good. In one of the pics that I circled twice, would you always want to take a horizontal section or is there a benefit to vertical sections? For the second pic with a singular section circled, when a section is this defined, would you simply pluck that to transfer? Plus, the dish labeled ISO has a very strong rhizo section I want to use but Im not 100% which way to come at it. I also have my bulk substrate ready to go which I culled from a few sources like JOC and Asura which was manure, straw, vermiculite, and a lil gypsum, pc'ed and cooled.

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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26593911 - 04/11/20 05:35 PM (3 years, 9 months ago) |
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So in the ISO dish with the two circles. Do you notice how it looks different from the 7 o'clock to the 12 o'clock region then it does from the 12 o'clock to the 7 o'clock region? I don't think it matters but it would seem like a rice shaped piece that's vertical would be best.
You're very close to a uniform looking isolate. I like the way it looks in the 7 o'clock to the 12 o'clock region i think anywhere in there is good.
They look great btw! Even that first PE one looks great. Now the second one you circled. You can try to cut, or use sterilized tweezers whatever you think will work. Truth is all that mycelium looks good. You can see how this rabbit hole gets bigger and bigger and the endless possibilities of tests to find something awesome.
But they all could be awesome with only small variations between them in flush sizes, potency etc..
Good job man!!
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26594089 - 04/11/20 07:11 PM (3 years, 9 months ago) |
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I really appreciate all the advice. Thanks for confirming that one section, I gravitated towards circling closer to the 12 o'clock position because it seemed thicker. Would that be better in hopes of stronger rhizo or go for more on the uniform side? The other PE plate I thought looked like it could grow out uniform. (honestly not 100% if you've hit iso when it's totally uniform? lol) Should I break up the breaks and let recolonize yet or do they all need to be 100% colonized first? I def see the proverbial rabbit hole, luckily I got plates and agar to try playing soon so I don't buy them all the time.
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26594118 - 04/11/20 07:26 PM (3 years, 9 months ago) |
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I don't think you'll need to break up and shake any of those bags. They seem aggressive enough they'll finish fine on their own I'm thinking.
Right uniform isn't necessarily isolated, not even sure if a pure isolation is even really a possibility genetically. It almost looks more aggressive near your thumb. But this maybe getting something you can see better than me in RL.
That's what I like about that other PE plate also is it might just stay that way.
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Roger Clemency
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26594168 - 04/11/20 08:02 PM (3 years, 9 months ago) |
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Nice looking plates. I never have tried to get a monoculture so no advice there but it looks like youâre using more parafilm than needed. Thatâs a personal preference thing of course but I watched a video that opened my eyes.
I pull out about 6 inches at a time and then cut lengthwise down the middle so you have a long flapping snake tongue basically, each side does 3 plates. You donât have to do that if you want the extra covering over the top and bottom of the plate but even so the film can stretch more than I see on your plates. Youâll notice lines on the paper and it doesnât seem like it but one section from line to line can wrap a plate.
Itâs probably a 2 inch section maybe? But if you try to stretch it straight out it will rip before covering the plate. Instead you get your piece on the plate, hold down with your thumb, and then turn your plate with that hand while guiding the film with your other. You have to use the bend of the plate to stretch the film, it kind of takes the pressure like a fishing rod.
You might want to try a practice plate or two because thereâs definite pressure involved and if it rips you get a massive movement of air and Oh Shit inside your SAB lol. You feed like a half inch or so and âturn-stretchâ it onto the plate, press it so it sticks, adjust your off hand back another half inch and keep repeating.
I hope this makes sense lol. Just a tip to save a little film cuz that stuffs a little expensive but 6000x nicer than using Saran Wrap or summat.
-------------------- Sour grapes, sweet revenge Heaven starts right where hell ends
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: Roger Clemency]
#26594233 - 04/11/20 08:39 PM (3 years, 9 months ago) |
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Quote:
The Abhorrent Chef said: Nice looking plates. I never have tried to get a monoculture so no advice there but it looks like youâre using more parafilm than needed. Thatâs a personal preference thing of course but I watched a video that opened my eyes.
I pull out about 6 inches at a time and then cut lengthwise down the middle so you have a long flapping snake tongue basically, each side does 3 plates. You donât have to do that if you want the extra covering over the top and bottom of the plate but even so the film can stretch more than I see on your plates. Youâll notice lines on the paper and it doesnât seem like it but one section from line to line can wrap a plate.
Itâs probably a 2 inch section maybe? But if you try to stretch it straight out it will rip before covering the plate. Instead you get your piece on the plate, hold down with your thumb, and then turn your plate with that hand while guiding the film with your other. You have to use the bend of the plate to stretch the film, it kind of takes the pressure like a fishing rod.
You might want to try a practice plate or two because thereâs definite pressure involved and if it rips you get a massive movement of air and Oh Shit inside your SAB lol. You feed like a half inch or so and âturn-stretchâ it onto the plate, press it so it sticks, adjust your off hand back another half inch and keep repeating.
I hope this makes sense lol. Just a tip to save a little film cuz that stuffs a little expensive but 6000x nicer than using Saran Wrap or summat.
Yes it does. The main reason, well two actually, but the strips sent with the poured plates I purchased were shorter or dry, and the area where it was only 1 layer was splitting and causing condensation and likely my previous contam issues. The film I bought I started folding the strip in half horizontally and then it still goes around each plate 2x. I'll def try that tip tho cuz your right, its pricey!
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26594565 - 04/11/20 11:03 PM (3 years, 9 months ago) |
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Quote:
rumfor69 said: I don't think you'll need to break up and shake any of those bags. They seem aggressive enough they'll finish fine on their own I'm thinking.
Right uniform isn't necessarily isolated, not even sure if a pure isolation is even really a possibility genetically. It almost looks more aggressive near your thumb. But this maybe getting something you can see better than me in RL.
That's what I like about that other PE plate also is it might just stay that way.
I meant to ask, do I need to dump the spawn bags into the substrate bags to colonize first? Because I also read about just mixing the two together in the monotub and letting it recolonize sub before casing so is it pref or pans/cubes like diff things?
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26595430 - 04/12/20 10:19 AM (3 years, 9 months ago) |
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It is all kind of a preference. BODs unmodified tub tek is so simple here.
I think stuff fruits better in tubs, more surface area, more set it and forget it operation. Cubes definitely seem to love tubs. I've never worked with Pan species so I'm afraid I'm not much help there.
Did you plan to follow a bucket tek for coir and verm?
Edited by rumfor69 (04/12/20 12:58 PM)
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26595696 - 04/12/20 12:37 PM (3 years, 9 months ago) |
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Thats pretty sweet, def low maintenance. Not sure if you had seen but I'm using those 32 QT Sterilite tubs, but I already modified them. I know its overkill but I'm trying to test some things before moving to larger space. I bought coir and verm but mainly followed a combo of things, but the substrate and pasteurization I mainly pulled from these: https://www.shroomery.org/forums/showflat.php/Number/25925194 https://www.shroomery.org/forums/showflat.php/Number/25987049 https://www.shroomery.org/forums/showflat.php/Number/26524100
Ive never done pans before either, but since they can share similar environments I figured do em both lol. My next test is going to be the RH/FAE since pans seem to like a lot. IM home so obviously the mist/fan is no problem but I have a modified ultrasonic I want to try dialing in that runs to all 4 tubs. I tested the flow and the humidifier actually regulates the moisture by increasing the air & flow rate, not the actual mist output so I'm thinking that will work in my favor (in theory lol).
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rumfor69
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Re: Need some advice for timing transfers and spotting contams? [Re: owntharabbithole]
#26595732 - 04/12/20 12:56 PM (3 years, 9 months ago) |
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I still cut 6 different 2" holes in my tubs and then I cover them with 3" 3M Micropore tape. For cubes at least in a tub with holes, they won't need injected air or humidity. Just tape the holes(or stuff with polyfill), mix up the tub, level it, snap the lid closed, and put under 12/12 light cycle. Then just wait till they're fruiting, don't have to do anything to them.
Here are two good coir bucket teks that A LOT of people use that work very well.
https://www.shroomery.org/forums/showflat.php/Number/24077162
https://www.shroomery.org/forums/showflat.php/Number/11916595
Edited by rumfor69 (04/12/20 01:08 PM)
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owntharabbithole
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Re: Need some advice for timing transfers and spotting contams? [Re: rumfor69]
#26616371 - 04/20/20 09:00 PM (3 years, 9 months ago) |
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So definitely getting what I asked for with learning and dialing new setups in lol I had spawned to bulk sub of manure/straw, verm, and some gypsum. Cased with peat/verm/calcium carbonite. Misted and placed back in original closet. Due to drastic temp flux I made the decision (with girls approval haha) to move into her office closet in the other room. I set it up the same which was 2 tubs stacked on each side with the humidifier between them along with a heating pad both hooked to Inkbird controller. Everything seemed good once set up, however got case of noob paranoia coupled with the threads reminding over and over you cant just crank it up and walk away so I kept checking every hour or so and went to bed. I woke up in the am to inkbird alarm on humidity because excessive condensation had pooled in the tube leading to the control tub and blocked the humidity and air flow. Im guessing lack of FAE, high humidity/temp, and/or a rushed setup introduced me to cobweb mold. And wow it IS impressively fast. The one tub went from small dime size to 3/4 of casing surface overnight and a second has small circular splotch. I treated with Hydrogen peroxide spray and seemed to clear it up. I fixed the condensation issue by using a space heater in the actual room and tweaked the humidity system based on JOC tek idea with central reservoir. I also got a Germ guardian air purifier that had $50 rebate:), UV, and Hepa to clean and add some flow. I first had it up on the top shelf on its side angled to blow downward towards the closet door and a small Vornado personal size fan to add some flow back up. Ive since scrapped that also and settled on current setup which is 2 tubs on the right, 1 on left (ultimately woke to the tub totally covered in light gray cobweb so I tossed it), I added a new tub I got on the floor on top of my seedling mat to bump up ambient humidity, put the purifier on top still blowing a door (around 150cfm so not much), I put the heating pad on the shelf with a 4" AC Infinity duct fan on top with few inches of duct hose on the front and back. Essentially pushing air down past tubs to back of purifier, towards door, and back up again. Since making these changes I saw huge difference in temp and rh stability. Ive been able to hold around 75-80 deg @ 90-97% RH with what appears to be great evap rate, possibly too good? Its very humid in there but there is no vapor droplets on walls and sub surface "looks" dry, but not sure. Im nervous to keep humidity on surface high because I dont want that particular tub to succumb to the cobweb again. Today I see some growth on this PE tub in question, but hard to say if its cobweb or mycelium because I do see white poking through in some spots. I was going to try carefully using peroxide again but not sure... any advice? Surprisingly, seems the pans Im trying are doing better that the cubes lol:-/
I also checked on the agar work and I think I finally got the plate, seems to have the right characteristics.
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