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Cookin' in the kitchen of love - Cybele's agar log


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#21 Asura

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Posted 24 September 2019 - 06:44 PM

 

Looking good!


I appreciate it bud! Is isolating wispy aggressive cultures specific to cyan’s or a general rule of thumb for all pan’s? All my pan cambo cultures are extremely tomentose. Picture the cotton on the end of a q-tip, but pristine, and glowing. It’s growing nearly a half-inch above the agar.

 

 

I can't speak to cambo, but several of us that have been growing cyans for awhile have independently verified that tomentose growth usually leads to overlay or mycelium that never leaves the vegetative state. I would try and fruit it. All pans are not the same. Might not be an issue for cambo.


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#22 cybele

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Posted 25 September 2019 - 01:38 PM

I can't speak to cambo, but several of us that have been growing cyans for awhile have independently verified that tomentose growth usually leads to overlay or mycelium that never leaves the vegetative state. I would try and fruit it. All pans are not the same. Might not be an issue for cambo.

I really appreciate the heads up. I have a close eye on my hard, and although the Cambo’s are all tomentose; each culture has differences. I’m going to try and fruit a tomentose Cyan culture as well. I like to see the right and wrong if that makes sense.

Edited by cybele, 25 September 2019 - 01:39 PM.


#23 cybele

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Posted 25 September 2019 - 02:14 PM

I figured today was as good a day as any. All cultures were inoculated two weeks ago. I have watched their characteristics as closely as possible, and I’m ready to make some transfers.

Pan Cyan, MEA
This culture gives three different options that are spread evenly enough about for my rookie hands to probably pull off. It seems that all three of these are wispy and growing evenly outwards. I am going to attempt to transfer a piece of mycelium from each to it’s own individual jar.
Note: on day 11 it started to grow nips. What ups with that? Lol!
E38258-FE-BE33-4313-8-B28-2-E5-F0261-DD9

Pan Cyan on PDA
Three different options here as well, but all are tomentose. Each differing greatly in appearance. Yet each growing at near the same rate. I will make one transfer from this plate to another in attempt to isolate a single strain with tomentose growth.
2-E04027-F-5687-48-B9-8-E45-1-C54-A8-EE9

Pan Cyan on MEA
Germination station right here. Several germination points on this culture. However, they are stacked together quite tight. I’m going to use my inoculation loop, and make five transfers from this jar. Many strains growing here look wispy.
EFFCBCC8-1-F76-4826-93-F1-167-D76365-FAD

Last but not least!
Pan Cambo on PDA. Germinated on day seven. When all three Cyan cultures germinated on day three. Four day lag, and faster grown than any of my cultures. Also growing up the jar wall. Two transfers from this jar.
13-E5-D8-EE-A702-4-EBE-A3-E5-0-CF4995042
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#24 cybele

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Posted 29 September 2019 - 08:27 PM

I didn’t get to the transfers until the 27th. I’m comfortable with my SAB, and it’s sturdy. It can just be a little cramped sometimes. So I decided to split the transfers up compared to doing all them in one round. I had to be tidier, and it was a little more time consuming. So I was only able to transfer half of what was planned for.

In a way that lead to some benefits. I get to critique my work before I complete my other transfers with these precious species. Because I felt really sloppy inside of the SAB during the transfers. I still need to modify the inoculation loop. I might go with a thicker gauge wire, and modify the shape. The one I purchased is flimsy compared to this resilient mycelium. I also felt sloppy two weeks ago with the same inoculation loop, and all of the inoculations were successful with no contamination. Which led to the successful inoculation of three pan cyan Alabama cultures.

I want to say, it’s truly crazy to start agar work and jump out of the gate wth pans.

So I placed one culture that was germinated into the SAB. Next to that I placed three new agar jars. I used a combination of the inoculation loop, and a scalpel. Trying my best to get the smallest piece of mycelium as possible.

We will see, as soon as I see something you will :)

#25 cybele

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Posted 05 November 2019 - 02:52 AM

Thirty seven more days have passed in this experiment since I last posted, and much has happened.
 

 

Being new to working with agar, and not knowing what all to look for I am keeping it small. That allows me to stay organized, but using these larger jars still has it's problems. With that being said I decided to shelf the Pan Cambo cultures as most appeared weak, and one never germinated. I plan to focus on one exotic species, and one cube species until I have an isolated, and coveted culture.

 

 

As with everything in this hobby one step forward, most likely means more questions! Most of my time spent will be gaining familiarity with the cultures on agar, and furthering the process of watching the culture's entire life cycle.

 

IMG-0870.jpg

There is a brown discoloration around the 12 o clock position of this jar that I am unsure of. It was one of the original cultures germinated from a spore print. I took multiple transfers from it, before the discoloration. At 55 days I expected more growth as this seemed to work halfway across the jar and just stopped. It may have to do with the thin pouring of MEA.

 

IMG-0871.jpg

Once again this is an instance of gaining familiarity. I do not believe that this is contamination, however it is easy to spot the discoloration. Can anyone speak on this culture?

 

 

IMG-0874.jpg

This culture is another from the original spore print. At 55 days this is the type of growth I envision. However, this jar was opened multiple times, and transfers were taken. Does this appear suspect to anyone?

 

IMG-0873.jpg

 

One thing I have noticed is that after about a week on the food colored PDY the cultures seem to make weird mountainous configurations then collapse, and turn green. Nearly 80% of my PDY cultures are not viable, unless I transfer fast.

 

I also started three new cultures of Golden Teachers so I can familiarize myself with cube myc, and get some decent genetics. I started with an old spore print.

 

Hope you all enjoy the post, and pictures!


Edited by cybele, 05 November 2019 - 02:53 AM.


#26 cybele

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Posted 14 November 2019 - 09:45 PM

It is definitely easy to spot some of the nasties. All three of the GT cultures had the same yellow bacteria within two days. So they went through a lengthy sterilization process, leaving me with no active cube species on agar. I didn’t expect much from a multi-year print that has literally been too many places. Including a storage unit for nine months.

Maybe I should have tried to germinate more plates. Either way I have two containers of GT consolidating, and then maybe we can snag a worthy clone during fruiting.

I took multiple transfers of the Cyans, and some picture worthy new growth should be here this weekend. Some of the less worthy cultures were discarded.

With that I was able to place some of the new lids on the jars. The others I inverted the metal lid to hopefully stop the suction problem.

I decided to stick with the MEA and activated charcoal until I figure out the food coloring issue.

#27 cybele

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Posted 20 November 2019 - 09:12 PM

IMG-1573.jpg

 

Fifteen total plates worked through for pans and I have only one specimen that I may further propagate. At this point of growth I cannot even tell if it is isolated. However, it is time to transfer it among three sterile jars so my only stock does not get ousted. Then it is time to test it. Even if this is an isolated cyan strain I lack anything to compare it to; background knowledge, nor another isolated culture. One will just not cut so I am going back to the print HARD.

 

For the cubes it is on to another round. As mentioned earlier my three plates from a GT spore print all failed. However, I have a tub pinning now of GT. I am going to snag some fresh, "clean" prints, and maybe if I am blessed a decent enough culture to clone.



#28 cybele

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Posted 22 November 2019 - 10:03 AM

I had some extra half-pint jars already sterilized, and even more in the dishwasher that I just purchased. So I decided to work some agar over this week before my Golden Teacher tub fruits. If I want to see success I’m going to have to put in the seat time. So instead of waiting for a clone I prepared some agar jars and stored them for when those tubs do fruit.

In the mean time I grabbed an extra twelve jars and decided to go head into the challenging prints again. I managed to streak six Koh Samui plates, and six Golden Teacher plates. If successful I’ll have started a KS culture, and diversified my GT culture collection (which severely needs it). If not, then I still have prints coming that will be much cleaner than my ancient stock.

Most valuable to me right now is the time spent working with the different tools necessary for the many procedures. Not to mention just getting comfortable inside the SAB.

Now we wait.

#29 cybele

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Posted 23 November 2019 - 12:20 AM

A4-F0-B222-FF89-4-DC1-802-F-F088-D49-D5-

87-B45-E86-6-FDE-448-F-B360-388-DB8-D35-

Pan Cyan on MEA?

#30 CatsAndBats

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Posted 23 November 2019 - 05:36 PM

I’m going to try a few different things. I feel like the condensation is going to my primary problem.

 

 

Store your jars upside down and drain/tap out the excess h2o when you inoculate.


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#31 cybele

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Posted 23 November 2019 - 08:14 PM

 

I’m going to try a few different things. I feel like the condensation is going to my primary problem.

 

 

Store your jars upside down and drain/tap out the excess h2o when you inoculate.

 

 

I appreciate it! I have already started draining it during inoculation. I am switching to the petri's as soon as my order comes in. All my posted cultures look free of contam's though?


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#32 CatsAndBats

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Posted 23 November 2019 - 08:25 PM

 

 

I’m going to try a few different things. I feel like the condensation is going to my primary problem.

 

 

Store your jars upside down and drain/tap out the excess h2o when you inoculate.

 

 

I appreciate it! I have already started draining it during inoculation. I am switching to the petri's as soon as my order comes in. All my posted cultures look free of contam's though?

 

 

 

 

As far as I can tell, but jars of agar aren't very photogenic in my experience.


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#33 cybele

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Posted 23 November 2019 - 09:20 PM

 

As far as I can tell, but jars of agar aren't very photogenic in my experience.

 

 

That is no fucking joke :laugh: I am seeing good numbers as far as contamination go. So I decided to jump into petri's. I waited way too long to work with agar, and I am not going to make the same mistake with equipment that I can learn from. Now I just have sit down and put together some safe cleaners. I have been using chemicals, and want to go "organic"


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#34 CatsAndBats

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Posted 23 November 2019 - 09:29 PM

 

 

As far as I can tell, but jars of agar aren't very photogenic in my experience.

 

 

That is no fucking joke :laugh: I am seeing good numbers as far as contamination go. So I decided to jump into petri's. I waited way too long to work with agar, and I am not going to make the same mistake with equipment that I can learn from. Now I just have sit down and put together some safe cleaners. I have been using chemicals, and want to go "organic"

 

 

 

Oh I got you on that subject.

 

5% acidity kills 99.9% bacteria and viruses, and I suspect that it also ruptures the cell walls of mold spores as well.

 

White vinegar is 5% acidity, but lacks any soap (read surfactant), so one could add a drop or two of dish soap/jet-dry, or one could just buy method anti-bac for surface sanitizing and Palmolive anti-bac dish soap for tubs and similar.

 

If one has access to a higher h2o2 concentration than one gets in the wound care section of the pharmacy (h2o2 at 3% is the brown bottle percentage), one can make a really effective sanitizing solution, but watch out, it's an oxidizer and will fuck metals and your skin up.


Edited by CatsAndBats, 23 November 2019 - 09:34 PM.

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#35 cybele

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Posted 24 November 2019 - 12:21 AM

 

 

 

As far as I can tell, but jars of agar aren't very photogenic in my experience.

 

 

That is no fucking joke :laugh: I am seeing good numbers as far as contamination go. So I decided to jump into petri's. I waited way too long to work with agar, and I am not going to make the same mistake with equipment that I can learn from. Now I just have sit down and put together some safe cleaners. I have been using chemicals, and want to go "organic"

 

 

 

Oh I got you on that subject.

 

5% acidity kills 99.9% bacteria and viruses, and I suspect that it also ruptures the cell walls of mold spores as well.

 

White vinegar is 5% acidity, but lacks any soap (read surfactant), so one could add a drop or two of dish soap/jet-dry, or one could just buy method anti-bac for surface sanitizing and Palmolive anti-bac dish soap for tubs and similar.

 

If one has access to a higher h2o2 concentration than one gets in the wound care section of the pharmacy (h2o2 at 3% is the brown bottle percentage), one can make a really effective sanitizing solution, but watch out, it's an oxidizer and will fuck metals and your skin up.

 

 

Just what everyone needs to know right there. For one Lysol/ Clorox wipes, Lysol anti-bacterial aerosol spray, and these other brands are expensive, period, much less in the amounts we use them. Or at least for myself. I am an overly tidy person by trade, so it surfaces in this hobby as well. For two, they are chemicals. Bad for the water, landfill plastic containers, and aerosol cans. Etc. Just bad.

 

So your telling me with some white vinegar at 10$ a gallon mixed at a 5/100 ratio kills bacteria and viruses? I thought I had read that somewhere, but was unsure if it also handled mold spores. Which for most is the real problem. I have had more mold, than bacterial issues. I am sure that changes person to person, and environment to environment. If you saw success at bringing down mold contaminants I will give it a round. I will add some dawn original.

 

I would not want to get into purchasing a commercial anti-bac, or using another chemical like anti-bac Palmolive. Ill try and keep it as simple as possible.

I greatly appreciate that!



#36 ethnobotanist420

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Posted 24 November 2019 - 02:07 AM

Awesome thread man! I’m about to whip up my first bit of agar here too... trying to do as much reading as possible lol

So your telling me with some white vinegar at 10$ a gallon mixed at a 5/100 ratio kills bacteria and viruses? I thought I had read that somewhere, but was unsure if it also handled mold spores....”


I think what Cat meant is to use the vinegar straight up... it’s at an acidity of 5% already since it’s 5% acetic acid. If you diluted it to 5:100 with water it would be (It's late and I’m high af so I’m not doing any math lol) way too weak.

Edited by ethnobotanist420, 24 November 2019 - 02:08 AM.

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#37 RutgerHauer

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Posted 24 November 2019 - 02:57 AM

Good log Cybele. You are doing great, learning while you practise.

 

I have not much constructive to bring in, but noticed your agar could be poured thinner, that'll save you some.

 

One other thing I learned myself about using MS solution on agar, that it would be beneficial to streak it out over the surface after you put a drop on it, in the same manner one would streak dry spores onto the agar. I realize that it would be harder to do with jars than petris, but maybe someone else can jump in here and tell if that is useful information in your case.

 

Stole this animated depiction of the technique from shroomery.

 

492818304-streak_agar_spore_solution.gif

 

Have yet to put this into practise myself, but reckon it's a good thing to try out.


Edited by RutgerHauer, 24 November 2019 - 02:58 AM.

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#38 Asura

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Posted 24 November 2019 - 04:08 AM

This is another great guide on agar if you are interested. It's a little overwhelming but

worth the read IMO. 


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#39 cybele

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Posted 24 November 2019 - 06:11 PM

Awesome thread man! I’m about to whip up my first bit of agar here too... trying to do as much reading as possible lol

So your telling me with some white vinegar at 10$ a gallon mixed at a 5/100 ratio kills bacteria and viruses? I thought I had read that somewhere, but was unsure if it also handled mold spores....”


I think what Cat meant is to use the vinegar straight up... it’s at an acidity of 5% already since it’s 5% acetic acid. If you diluted it to 5:100 with water it would be (It's late and I’m high af so I’m not doing any math lol) way too weak.


Hahahaha thanks for the clarification! It was late, and I was high af as well. A gallon of vinegar will still be remarkably cheaper than my current cleaning product line up. I started all of my agar work the first night after I completed a four month contract. (I don’t smoke while contracted). My S/O thought it would be a great idea for me to do some concentrated oil, and everything was fine until I sat down in my SAB. I was blitzed. Surprisingly only had 3/24 jars contaminate. Tonight’s my last night of smoking because I’m about to contract overseas. So the past few nights I have been knocking out myco work, listening to hip hop too damn loud, and getting way higher than usual.

What else are you supposed to do with three months off lol!

#40 cybele

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Posted 24 November 2019 - 06:17 PM


Good log Cybele. You are doing great, learning while you practise.

I have not much constructive to bring in, but noticed your agar could be poured thinner, that'll save you some.

One other thing I learned myself about using MS solution on agar, that it would be beneficial to streak it out over the surface after you put a drop on it, in the same manner one would streak dry spores onto the agar. I realize that it would be harder to do with jars than petris, but maybe someone else can jump in here and tell if that is useful information in your case.

Stole this animated depiction of the technique from shroomery.

Have yet to put this into practise myself, but reckon it's a good thing to try out.

I deem myself kind of obsessive with all amounts of research. There is just so much in this world that is fascinating. Yet there is a reason experience is the best teacher is a long standing quote. It’s true, and always will be. No matter how much I research the time in the “field” is always more rewarding. I hate that it took me so long to persuade myself to come to afar but here I am!

I streak my plates with various symbols during spore transfers, and love watching the young translucent hyphae rip across my symbols. I definitely need to pour my jars thinner. I’m moving to plates though so I’ll have to adjust for them.

Edited by cybele, 24 November 2019 - 06:24 PM.





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