How to salvage a contaminated petri dish while protecting your lab

[@Whatsittous]

Never clicked on a video so fast in my life

[@themyceliumnetwork]

LOL

[@Pheno-Hunter]

Same 😂

[@BETTERthan_Yesterday]

Lmao same! I was literally watching a different video and ditched that one for Gary bc I trust him wayyyyy more.

[@slimgrows]

Hey buddy just wanted to say thanks...you willy myco and 90 second mycology have taught me everything and been a huge part of me setting up my home lab... 🌲 ❤️ 🌲 🍄 💜 🍄

[@jdmarsee]

Same here dude… I watch a ton of their vids

[@themyceliumnetwork]

I used to work at "Nature Lion Mushrooms" the entire place was one big mold and bacteria factory. my home lab got contaminated from working there & it took me months to clean all the molds & bacteria out of my home lab. work smart in your lab folks :) its not worth the months of cleaning & scrubbing.

[@patnowhere]

I don't know how the capital EFF I wasn't subscribed been watching for over a year.. better late than never! Thank you for all you do Gary and congratulations to you and your family!!

[@front7inexfatal432]

Miss the videos Gary, good to hear from you again. Happy holidays to your family brother

[@DeShark88]

I searched for this exact video (ideally from your channel as well, since you always do it "the proper way" and you provide the additional background knowledge too) only a couple of weeks ago. Can't believe I didn't find this! I had some nasty-looking pink stuff growing, from what I think was a contaminated spore syringe (E-coli?!) I just kind of winged it and somehow all went well (or so I think up til now). I really couldn't find much about isolating good mycelium from contamination, so it was a total surprise to me that it worked as well as it did. My worry now is that I've isolated a random mold rather than mycelium - what's the best way of knowing? It's white and it's mycelium-like, but I don't want to inoculate a jar of grain spawn and wait months only to discover I've grown a ton of some yeast strain or something 😂

[@skyelord6229]

Congratulations Gary and Addie!

[@bounceurabdelaziz4973]

Thank you Gary, great video, my greetings to you from Algeria🇩🇿.

[@SamsLiberty]

Congrats on the new baby!

[@JDVokal94]

Congratulations on the new edition homie!! Mush Love!!🙏🏾💯

[@SpaDeKo]

Crazy timing. I just poured some agar "plates" in preparation for some king oyster mycelium with some bacterial growth. Luckily the mycelium is easily outgrowing the bacteria, so I should have some clean areas for transfer. Thing is, I don't have a pressure cooker, so all I do is pasteurize(And barely that), and my SAB is just a rather big plastic bag. I still have a 10-15% success rate even at this low-tech of a tek. Most people might get frustrated at these low percentages, but as a hobby, I'm perfectly happy with my few successes! :P

[@bradzimmer239]

Could you use a few drops of peroxcide to contain the mold spores? I had some Trich mold in a grow bucket, where I sprayed and then removed that infected area with an alcohol cleaned spoon. Just an idea.

[@jjohnston73]

I had the same question.

[@MaxParadiz]

I really like this video emphasizes examples of contamination that you, a well-seasoned expert with a flow-hood, experience during your own work. Often videos focus on those plates that went well rather than on those that went wrong, and as a beginner it made me over-estimate the success that people had with their plates. I would get maybe 3 or 4 clean plates out of 10 and I would be very disappointed. I would spend a lot of time trying to "optimize" my technique and figuring out why it went so wrong. Then I realized than even with 40% success rate I would end up with more plates than I needed... Now I don't worry so much about this, I just pour more than enough plates assuming things will go wrong. I think accepting that contamination is part of the game is as valuable or even more valuable than optimizing the conditions to get rid of it :-)

[@PlasmaOne]

That induction heater looks mighty useful for SAB work since you don't need fresh oxygen to use it. I always torch too much inside my SAB and run out of oxygen.

[@jcondron]

Can you point me to a source that might help me identify contam versus mycelium on agar/ Perhaps with pictures? Lots of pictures?

[@MarianaDee22]

Thank you so much. This was very helpful!!!

[@haiku859]

Appreciation ~ Very informative channel Keep em coming 🙏

[@RustyColonsCPA]

Thanks Gary. Great info, as always!!!

[@paulmaestas741]

Aren’t you moving the spores around by moving it like that or does it take more then that for it to release spores?

[@Lemonz1989]

Thanks for showing us how you do it ☺️ Just wondering; wouldn’t it be easier (and cheaper) to use a reusable inoculation loop to quickly scrape a piece of mycelium off of the dish and transfer it to another dish? Also, I would be very interested to see if you could make videos testing UV-C light as a method to reduce risk of contamination. Perhaps to see if you could sterilize spore prints before use with a quick burst of UV-C, without killing/inactivating the spores themselves. And perhaps give a spore syringe a dose of light after you’ve made it, to kill potential contaminants inside the syringe without destroying the spores in the process. It would also be nice to see if you could sterilize Petri dishes after pouring, to stop any contamination from taking hold. This would make things so much easier for us who don’t have laminar flow hoods. I know some labs use short bursts of UV-C on mycelium to induce mutagenesis, to quickly develop new phenotypes for research purposes. Just a side not to anyone interested in working with UV-C light. Don’t look at it, and don’t expose your skin to it. It can cause damage to your eyes and sunburn-like damage to your skin that can cause cancer.

[@risottoingiallo3448]

Hi, I have a question, I'm trying to grow in lions mane agar agar, the recipe for nutritious agar that I used is this: 500 grams of water, 10 grams of agar agar gel and 15 grams of honey. and the mycelium of the lion's mane is very slow and always remains in the same place while the original sample continues to grow, forming the typical hair of the mother mushroom. what could I do? Did I do something wrong? ah, I'm keeping it at a temperature of 24.5 degrees celsius

[@syberphish]

Hey man congrats to you both on the kiddo! ^__^

[@1ntwndrboy198]

To avoid contamination to your lab couldn't you have a hepa filter vacuum sucking in air right when and where you open lid? Then when open smother the contam with vasoline dab? Curious if this would work 👍🤟✌️

[@FreshfromtheFarmFungi]

I wouldn’t risk it but maybe just as a fun side experiment it might work

[@jdmarsee]

I love your vids dude….love the shirt!!!

[@jasonmillar2626]

from my experience contamination on the edges of the dish is generally caused by the laminar flow hood, either not running the flow hood for long enough before using it or not wiping it down before working Infront of it... also should mention that unless you have a "clean room" you hardly changing the spore count in that room at all. Your probably got more spores on your cap than in that petri dish

[@FreshfromtheFarmFungi]

a good hypothesis to be tested 👍

[@dhaval4570]

Lovely video, quite helpful! As another guy said never clicked faster 😂

[@konstantinNeo]

How about pouring hot agar on top of the gelled agar with contamination? Isnt the mycelium grows to the top and the the contamination stays burried?

[@FreshfromtheFarmFungi]

it can work I think there might need to be antibiotics added in this case or some organism might survive still. I have not tried it though it might be very effective

[@meachyofficial]

Could you have used an inoculation loop to scrape the mycelium instead of cutting the agar?

[@FreshfromtheFarmFungi]

yes

[@Frittierer.111]

I am getting up my third and fourth back up plate at home. But I am never really sure when to transfer.. Should I wait until the mycelium looks strong enough or should I as soon as possible cut it out, to not let the already contaminated plate spread even more... really difficult to decide?! Any suggestions? Greetings, great Video as always!

[@FreshfromtheFarmFungi]

I like to wait until the mycelium is about 80% grown out - you can see any contaminants on the underside at this stage and try to avoid it on the surface side if that makes sense and the leading edge is always the cleanest

[@DangerRussDayZ6533]

Hey Gary, you should check out the mycocoil induction heater.. it has a foot pedal so totally hands free if needed

[@FreshfromtheFarmFungi]

thanks! After I got this one MycoGeeky made one with a pedal as well - going to put the scalpel mileage on this one before upgrading 🙏🏻🍄❤️

[@DangerRussDayZ6533]

@@s7r49 Can I ask why? I don't have an induction coil (yet) but it's on my list. I was definitely leaning towards foot pedal, so I'm interested to hear why someone would recommend against it.

[@MrOiam]

Could you recommend a book or texts on separating contamination working with dishes etc

[@FreshfromtheFarmFungi]

I like “Radical Mycology” by peter McCoy - he goes thoroughly into contams. Also Paul Stamets books - check out this list www.amazon.com/shop/influencer-a88d1601/list/3KAE5ZYPR7PAX?ref_=aipsflist_aipsfinfluencer-a88d1601

[@MrOiam]

@@FreshfromtheFarmFungi will do ,,,thanks

[@ineshianewton7740]

Hello could you tell me how to clean a monotub that had trichoderma?

[@FreshfromtheFarmFungi]

I recommend soaking in a bleach solution for at least 20 min (I usually use a larger bin filled with 10% bleach and soak overnight) then dry off and wipe with isopropyl alcohol you will be good to go 👍

[@ineshianewton7740]

@@FreshfromtheFarmFungi ok thank you...and will the bleach give instructions to make 10% solution?

[@xojioghbiu]

Thanks, but what to do if transfers are not the option anymore? Could gentamicide or anything help in bacterial contamination?

[@FreshfromtheFarmFungi]

it can help knock it back but it would have to be added to the agar aseptically while it was being prepped. I steer away from antibiotics unless im working with outdoor spores or difficult tissue since it ends up in the environment at some point and can lead to antibiotic resistance. It also can become a “crutch” that cultivators lean on instead of learning proper technique and add costs when you shouldn’t need it if that makes sense.

[@FreshfromtheFarmFungi]

also, look into water agar it’s safer for the environment and can do similar things

[@xojioghbiu]

@@FreshfromtheFarmFungi Thank you!

[@YungStevenSeagal]

what the bag @14:39?

[@MikeNaples]

Breakfast cereal for his pet mycelium.

[@minivanvibesinc]

What is that device u use for sterilization of the blades called?

[@FreshfromtheFarmFungi]

it’s an induction sterilizer by @mycogeeky

[@patricksweetman3285]

Great work Gary. Do you have a selective medium for basidiomycetes? I tried to come up with one 18 months ago. It had ampicillin, carbendazim, and a phenol, guaiacol, but I couldn't suppress the penicilliums. I tried adjusting the pH and salt concentrations but gave up when nothing worked.

[@FreshfromtheFarmFungi]

I don’t have an isolation media for basidiomycetes but I do like DRBC - it slows growth enough that you can weed out penecillium 👍 just gotta check daily

[@kristhomas6474]

Dude im just a beginner and havent even been able to get my first flush, but you are stating how careful you need to be when openning the agar plate and take a piece farthest away and you dont even want to use your laminar flow but then you are shaking the contaminated agar plate and dropping in on the table. Like i said I'm new to this, but wouldnt those action create the possibility where spores from the contamination dislodge and contaminate the whole inside of the dish. Im just learning and trying to find best techniques and practices

[@FreshfromtheFarmFungi]

for this particular case, the biggest factor for contamination is the surface moisture (yeast don’t have spores and it’s rare for bacteria to become airborne in a dish) for mold this is a very big concern. Also, these videos were filmed for teaching I usually am more focused when doing work for products - I will revisit these topics in the future now that my video making skills are improved.

[@fishmut]

From watching this I thought about practicing the same as I do when I take cuttings from plants to propagate , always always do more than you need , never ever just do one , there is a strong chance some won’t make it, so if I did say 5 and 2 are contaminated, I won’t have a problem disposing the contaminated ones at all , not worth my time , if you still have the mother for your samples start again if needed , much more easier than fiddling with the sick ones , but like if that’s all you got (1) than have fun lol. 👍

[@jeremylastname873]

It’s not “one single spore” of penicillium. 🤓 Thank you for your endeavor to educate and good luck with your family and career.

[@zensuufu]

I thought that was funny too, but to be fair he meant that it was inoculated with a single spore.

[@FreshfromtheFarmFungi]

yes I meant one “colony” but didn’t realize until after hah good catch! I one take these videos to keep them as raw as possible and nothing slips by! 🙏🏻🍄❤️

[@gamephly]

Do petri dishes with vented lids increase the risk of contamination?

[@FreshfromtheFarmFungi]

they shouldn’t if you use parafilm

[@LarsLarsen77]

News flash: your lab is already full of mold spores, because all air is.

[@Mushmammy]

Dammit. I thought you meant a link on how to MAKE the sterilizer 😂

[@FreshfromtheFarmFungi]

there are some good videos out there on DIY barrel steamers and horse trough ones

[@InconspicuousOwl]

🔥 🍄 💉 🧤 🚀 💩

[@mikeg5616]

I keep getting a milky type of contamination on the surface of my plates. Wtf is it?

[@terrymckensey6055]

Bacteria i believe