Importance of quality control for liquid cultures: dealing with contamination in LC mushroom culture

[@tylercunningham1878]

Gary, you have no idea how ecstatic and thrilled I was when I saw you put the beginning and the results. Some you tubers would split videos and it’s better personally when we can see it in the same video. Stellar job with these new breeding cultures

[@johnathanfaircloth9639]

It's good to see that the only ones messed up had imperfections in their Jars.

[@jerodyoumans6002]

One of the better videos in/or on mycology before, after and in betweens, with examples of mycelium growths including names and images.Thx!!

[@pauloantunes8826]

Gary!!!! I just made my first LC! Thank you a lot for your techings!

[@Sssanbo]

LC requires being super sterile. I remember being worried that i was contaminating the LC every time i even put the needle in, even when testing.

[@ryanennis3199]

Same haha

[@shortchanged.]

Yep

[@TheGoodFunGuy]

Very informative vid, Gary! Thanks!

[@taylormadefungi9004]

I fill the void on bottom side for those grey injection ports with RTV so they last a little bit longer

[@AaronAlso]

They are too cheap to worry about.... don't be an LC tipping barbarian and install a simple female luer lock to 1/8 barb fitting with a bit of silicone tubing (RTV and basic DIY required) Use a male luer lock plug to seal the lid. You can install the injection port, but will rarely use it. Just remove the plug, lock on syringe

[@taylormadefungi9004]

Right on man looking forward to getting some of your new strains

[@ClownWhisper]

I spray everything with peroxide then alcohol wp mins later. I have been infection free for more than 5 years

[@boomingtrendz]

Hey, what’s up Gary very good informative video on liquid culture. Thanks for sharing this I do have a question what size are those ball mason jars? Thanks again for all you do in our community.

[@FreshfromtheFarmFungi]

these ones were 1/2pints from walmart

[@ShawnTheRazor]

I purchased your Pink Oyster Rosebud LC and have been very happy with it. I haven't fruited it yet, but just enjoying watching it colonize grains, agar, and substrate currently. This video makes me wanna pour some black agar.

[@FreshfromtheFarmFungi]

awesome! love to hear it 🍄❤️

[@unclebobsbees4899]

Thank you, this was informative. Especially for the new kids, like me.😊

[@wajahathusain1175]

Please tell rapid qc detection for mycelium culture

[@user-xb1ht4py2v]

great video. I have a few questiosn though. Is the purpose of puling ot a syringe full of LC then squirting it back in too mix up the culture? AT time 7:44 when you tip the jar over to fill the syringe alot of liquid drips out onto the table; why, what, and from where?

[@FreshfromtheFarmFungi]

it’s just to demonstrate the act of pulling - normally I inoculate grains right away. The liquid was alcohol from the injection port being sprayed

[@user-xb1ht4py2v]

@@FreshfromtheFarmFungi Thanks. that also answers another question i had. I use lots of alcohol and usually items stay wet until it evaporates. To see yours wet enough to drip lets me know I am not using too much. Got my first batch of LC going now. Thanks a ton.

[@CMAenergy]

I made a simple flow chamber But what is the best sized filter to use How many microns I think this one has 2 m Also on a Petri dish When there is contamination and you see some good spores . Is it possible to remove some good spores and see if they will produce without contamination I assume not but just a thought for a beginner

[@Andrew.84s]

Keep putting those videos out bro🥳🍄🍄🍄🍄

[@Thomas.V.-223]

Thanks man for the tips and tricks...

[@jerichojames4769]

does every withdrawal of liquid culture have the possibility of contaminating the entire liquid culture?

[@FreshfromtheFarmFungi]

it has the possibility yes but if you use good protocol it is very small risk. Make sure you wipe with isopropyl and use aseptic techniques

[@johnriperti3127]

Fascinating Gary, thanks!

[@buself]

I have been constantly getting trichoderma on all my LC attempts.. My pressure cooker is only at 13 PSI, do you think that could be the issue? I sterilize my agar and LC for about 20 mins. Also I tried making agar plates(which is sterilize the same as my LCs) and left them without inoculating for few days - no green mold is growing. I am really confused and frustrated so far.

[@thejohn6614]

Are you flame sterilizing your syringes when you're going into the lc? What do you have for a filter on the lc jar?

[@koltoncrane3099]

If you’re going to test your LC on agar, and grow them to ensure they’re clean and strong why then do you use little bottles? Wouldn’t it cut down or reduce your labor if you say used gallon jars and then did quality control tests on that gallon jar? That way you ensure that one jar is clean and good but then because it’s say a gallon you can get way more out of it. The expense per jar would be lower. In accounting they talk about fixed and variable costs. In this you could reduce your labor costs a lot per LC syringe you sell by going to large jars to reduce quality control costs

[@FreshfromtheFarmFungi]

yes - mostly because of space issues and because I like to have fresh LC but at our new spot we will be scaling all of this thanks for the suggestion your head is in the right spot! 🙏🏻🍄👍

[@kevdogg3606]

If you have a syringe filter on your lid...do you leave the lid tight or back it off before placing it in the pressure cooker ?? Thanks for your time..

[@FreshfromtheFarmFungi]

I back it off a quarter turn just because it may loosen the seal

[@buself]

hey, everyone says that you should store your agar plates upside down, but seems you don't do it. is it really needed?

[@FreshfromtheFarmFungi]

long term yes to prevent condensation from forming a pool on the surface and choking the mycelium. For active growth it is best practice but isn’t going to make or break it. I do store them agar side up refrigerated for this reason.

[@BadgerLaser]

interesting video many questions answered and lots to think about. what is the idea with pumping the syringe several times please ?

[@FreshfromtheFarmFungi]

it just primes the pump and mixes the culture so it’s more homogeneous

[@ClownWhisper]

And I got cultures that are totally foggy and are clean as a whistle although I do use nutritional yeast which can fog things up to

[@ystes23]

Do you put parafilm in the petri dishes after inoculating?

[@FredHam-p5s]

I'm having problems with contaminated lc. Who has the best golden teacher lc

[@CharvelJackson]

Tks Gary, I think there are some of us with no flow hoods, so, this testing step introduces another vector that is independent of the original LC and seems like a way to get some false positives for contains which may not be there? I guess grain in a jar would be the best alternative for us non flowhooders?

[@FreshfromtheFarmFungi]

no, you should still test, just sterilize the needle and injection ports right before testing (flame the needle and use iso on the port)

[@CharvelJackson]

@@FreshfromtheFarmFungi Thanks for your time, I am not understanding how flaming and spraying needle and port when getting my LC out helps what seems like and Open aGar Dish when I open the lid to put LC in? I wont know if it was in the LC or if it came from the environment because I dont have sterile environment/lab. In grains I have the environment sterilized and the LC doesnt ever get exposed. ??

[@spencerreiser7765]

Buy sterilized agar plates or make your own flow hood (SEE Also gordotek)

[@AndrewPak-kt6qy]

Once you've confirmed the viability of a culture, can you then use the same syringe you used to test? I want to reduce the number of times I have to poke into the jar and use less syringes. Thanks for all the info!

[@FreshfromtheFarmFungi]

yes you can do that just do few ml’s each and there should be enough for both

[@AndrewPak-kt6qy]

@@FreshfromtheFarmFungi Awesome, appreciate the reply and all the great vids!

[@blahzes]

Wouldn't it be cheaper to incoulate small grain jars?

[@FreshfromtheFarmFungi]

yes but contaminants can hide in the 3D nature of a grain jar when plates filter out this dimension

[@trevorbraden5448]

What brand lids are those ?

[@sansulabrahma349]

How many temperatures store

[@jamesrapkins4935]

Black Charcoal Agar! :P i must have this! thanks Gary! MUSHLOVE , will be eating your Turtle shell Beech in a week. Colorado oyster was deeelicious ty x

[@eXWoLL]

Is activated charcoal agar really good for QC? Because from what I've read about it, it's supposed to reduce/elminate bacterial growth, in which case would kinda of neglect its usefulness in QC since if its effective in its purpose, you wouldn't see any bacterial growth from a contaminated LC.

[@FreshfromtheFarmFungi]

From my experience black charcoal isn’t antimicrobial, it may slow the growth which would make it easier to discern different colonies before they all grow and merge into a larger biofilm but mostly it stabilizes the pH in the media and can help absorb some of the exudates from the mycelium to prolong the life

[@Foxxxxx96]

@@FreshfromtheFarmFungi very interesting

[@kicknadeadcat]

I used honey and peptone 15 psi 30 mins and the LC was a little cloudy right out of the cooker. I have used honey alone and maple syrup alone and it was clear. Could it just be peptone particles? Mycelium seems to be growing.

[@FreshfromtheFarmFungi]

possibly - need to test on agar

[@littlerhino2006]

Okay what are the likely consequences of a home grower skipping this step, is the mycelium able to consume the bacteria or will the resulting mushrooms all be contaminated?

[@FreshfromtheFarmFungi]

it will probably cause contamination problems and be discouraging

[@MayhemKeys]

When isolating or germinating spores, do you ever use antibiotics?

[@FreshfromtheFarmFungi]

I don’t but it’s a good last resort

[@chadgurney1672]

Where's a place to get liquid culture testing supplies

[@FreshfromtheFarmFungi]

amzn.to/331iD1c here these work well

[@chadgurney1672]

@@FreshfromtheFarmFungi thank's love the videos

[@raytry69]

Tip: put your mic away from the flowhood, that will reduce noise.

[@brianbordenkircher52]

Thank you for the info!

[@nickfranco2259]

Why did the lc drip out while you where filling your syringe??

[@FreshfromtheFarmFungi]

the lid wasn’t on tight enough

[@Nope-hz1ux]

@@FreshfromtheFarmFungi it came from the filter when you were pumping the syringe back in. why are you pumping it like that anyway?

[@antoniosa932]

@@Nope-hz1ux i saw that too, never seen that before

[@zeuscaboose7202]

How do you decon your plates for another use?

[@FreshfromtheFarmFungi]

We only use them with agar once, but then for spore printing we wipe them clean and spray down with isopropyl alcohol

[@Basket_Knot]

You look so much like a guitarist named Adam Demicco!

[@FreshfromtheFarmFungi]

neat! I used to play the trombone and bass guitar in high school so I appreciate the art

[@judyausmus6130]

Like your video

[@jschultz353]

have I been mispronouncing agar this whole time?

[@FreshfromtheFarmFungi]

people say that I have - midwesterners say “Aygar”. Upstate Ny’ers day “Ahgar” it’s all the same 🤷‍♂️

[@williamandriusaitis2397]

You should wear a gown and a mask