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#21 rockyfungus

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Posted 17 December 2020 - 11:32 AM

I'm clumsy and move way too fast. Broke a few cleaning, broke a few innoculating, broke a few going up and down the stairs...I should of known better based on me being a complete spaz.

Now I have 3 glass tops and no bottom part to fill with agar. Trying to brainstorm a way to use it still.

Sorry for derailing you, Spahro.


Edited by rockyfungus, 17 December 2020 - 11:33 AM.


#22 Saphroziac

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Posted 18 December 2020 - 03:16 PM

Here are some of the results from my agar transfers. A few were inoculated on 12/13, and others a few days earlier. So far it looks as if no contaminants came through on 9/12 jars, when I opened the jar in front of the flow hood, as I perfect my quick sterile preparations and procedures.

 

1.jpg 2.jpg 3.jpg 4.jpg 5.jpg 6.jpg 7.jpg 8.jpg 9.jpg


Edited by Saphroziac, 18 December 2020 - 03:21 PM.


#23 jrh

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Posted 18 December 2020 - 03:39 PM

I'm curious what grain you're using to spawn. I use whole grain oats with the hull, and those look like the few oats that slip free from the hull during cooking.



#24 Saphroziac

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Posted 18 December 2020 - 03:42 PM

They're Rye Berries and there is also a bit of clumped up Gypsum in there. :D


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#25 jrh

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Posted 18 December 2020 - 03:43 PM

I guess a grain looks like a grain looks like a grain. :)



#26 Saphroziac

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Posted 23 December 2020 - 05:19 PM

Hey guys, here are some Rye Berry Quart Jars I transferred over from some Agar. on  12/3, 12/13, 12/15, and 12/17. It feel like they are going super slow, it has been only ten days roughly. They are all in incubators at 70-80-F.

 

This Treasure Coast Jar is doing the best out of all of them. I tossed a bunch of Gypsum in there, and it all collected at the bottom.

2.jpg

 

These Treasure Coasts was inoculated a month ago on 12/3. They look Vastly different than the previous Teasure Coast; they seem a little weaker/thinner or something.

7.jpg  8.jpg

 

Most of these Rye Berry Quart Jars are barely taking hold still.

9.jpg 6.jpg 5.jpg 4.jpg 3.jpg 1.jpg

 

These BRF Jars are killing it though.

 

Jalisco, took 2 weeks just for this guy to appear.

10.jpg

 

Tampanensis

11.jpg 13.jpg 15.jpg

 

Malabar

12.jpg 16.jpg

 

Pan Cyan Australia

14.jpg

 

Gallindoi

17.jpg

 

 

I feel like I should just do BRF Jars for now on instead of Rye Berry Quarts. :/  :mellow:  :dry:


Edited by Saphroziac, 23 December 2020 - 05:20 PM.

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#27 jrh

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Posted 23 December 2020 - 07:45 PM

I wonder if those 2nd set of TC jars aren't trich or something. That doesn't look like strong, white cubensis mycelium to me.



#28 Saphroziac

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Posted 23 December 2020 - 08:10 PM

It's looked that way for quite some time, even after shaking. Not sure what to think about it. Might try and get it on Agar again to see what it does.



#29 Saphroziac

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Posted 28 December 2020 - 07:27 PM

Does anyone know what these contams are? Are these bacteria? I am pretty sure these are bacteria, they had the super gross smell in them. Crap. Someone told me I needed to leave the weight off my PC for about 15 minutes while it's heating up to remove any air pockets that might form inside of it halting it from getting to true PSI 15. So I went wrong in either that regard or my Agar plates I chose were in fact bacteria contaminated and I didn't see it. (But the plates looked clean! :(....)  Wow there is even a odd mold in one. Any suggestions appreciated. <3

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Edited by Saphroziac, 28 December 2020 - 08:11 PM.


#30 Saphroziac

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Posted 28 December 2020 - 11:44 PM

What is in this Petri Dish? Is it Pink Mold?

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#31 Auhron

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Posted 29 December 2020 - 01:27 AM

Slimey evilness. Kill it with fire!

 

If that Petri Dish contained Potato Dextrose Agar (PDA) then it is most likely Clostridium Puniceum, a bacteria that presents as pink slime and infects potatoes. The endospores are thermophilic (heat resistant), as such unless germinated before the sterilization cycle they can survive the PC or Boiling.

 

The pink slime could also be the common household pink slime Serratia Marcescens. It usually presents more reddish than pink, and normally grows where standing water is likely: bathrooms / kitchens. 

 

It could also be Lycogala Epidendrum, an amoeba that can look like a tiny pink puffball, turning brownish with age. The other two options seem more likely.

 

Kill it.  


Edited by Auhron, 29 December 2020 - 01:35 AM.

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#32 Saphroziac

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Posted 29 December 2020 - 02:45 AM

I did try using PDA and these petri dishes might be one of them,  this pink mold is in 5 petri dishes, all with my AA+ Syringe. None others. So it could be coming from the syringe, or from the PDA. I probably wont do PDA again.

Where do you find out about al these different molds?



#33 Auhron

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Posted 29 December 2020 - 03:17 AM

It's not the PDA's fault if it was Clostridium Puniceum, just the potatoes. Adjust the procedure so you grind/mash/puree the potato the night before and soak it, then proceed as usual. Similar to grain prep, the spores should germinate overnight then die during the sterilization run.

If it was the syringe, well that's shitty and I hope you got at least enough healthy growth somewhere that you can make transfers.

 

Google. trying different search terms. cross referencing ones that meet the description against multiple sources. wikipedia can be a powerful tool as well. and microbiologyresearch.org among other similar sites. Narrowing down the search to weed out the irrelevant information is the tricky part, but the answers are out there.  

 

learning how to use advanced search features is a game changer. Just the basic 4 really:    " ",   -,   +,   |   

http://booleanblackb...ers-explained/ 

Ironically the best instructions to using search engines don't come from google. That would limit your exposure to their revenue machine.

 

https://www.informit...e.aspx?p=675274 Is another decent one. explanations are worded better.


Edited by Auhron, 29 December 2020 - 03:21 AM.

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#34 jrh

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Posted 29 December 2020 - 07:37 AM

The pink slime could also be the common household pink slime Serratia Marcescens. It usually presents more reddish than pink, and normally grows where standing water is likely: bathrooms / kitchens. 

 

OMG I have this everywhere in the house. I need to look it up. It shows up anyplace that is constantly wet -- toilets, sinks, the bottom of my toothbrushing cup, pet fountains and filters. THANKS for identifying this.



#35 Saphroziac

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Posted 29 December 2020 - 05:01 PM

Heres a couple more inoculated Rye Berry quart jars from agar. I've got Stargazer B+ Golden Teacher, and Treasure Coast. Most of these are excruciatingly slow, but stargazer seems to be doing alright. I hope no wet spots/bacteria appear on these. :(

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#36 Saphroziac

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Posted 08 January 2021 - 04:44 PM

Can anyone recommend the best way to go from fully colonized BRF jar to agar? or BRF to Quart Jar?


Edited by Saphroziac, 08 January 2021 - 04:44 PM.


#37 newmoon

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Posted 08 January 2021 - 05:22 PM

Open the jar in aseptic conditions and scrape a small amount of mycelium onto a scalpel or inoculation loop, and then place that on the agar plate.

 

To go to a jar of grain you can use TVCasualty's slurry technique.






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