The New Inoculation Lab
So, uh, right now we're in the laboratory. This is actually, uh, a newer lab that I put together, and I'm primarily utilizing it for mostly spawn and culture type production, so this way I can kind of take some of the load off the other lab, which is mainly used for a lot of the inoculation of the sterile sawdust and bulk substrates, and kind of just do more and be able to be more ahead.
So today we've, uh, sterilized some liquid media for liquid culture inoculation, so it's actually been able to cool right in front of the sterile airflow of the HEPA filters, and at this point we can unload it and begin conducting the tissue transfers of some freshly colonized agar and grow out our mycelium like that. [Applause]
So we'll just depressurize the chambers, and that just escapes any built up pressure inside of our sterilizers. So, yeah, in these jars we have 500 milliliters of our liquid media. So it's just a nutritive liquid broth that we can use to expand out the mycelium, and each jar is equipped with a magnetic stir bar, of course, so we can basically use that to agitate the mycelium and break it up, allowing it to be intake through a syringe.
So I'm just gonna pre-uh pre-label the jars before I get started. That way, that helps with, uh, just making sure everything's labeled. The worst thing that can happen, I guess, is inoculating one of these and not having it labeled properly, only to find out several weeks later that it was the strain that you didn't want, so, or that wasn't intended, and that could be for your, you know, a certain substrate preparation or a certain recipe pertaining to a certain strain, and you just didn't get it right because you just didn't know.
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Southwest Mushrooms Genetic Library
The strains themselves, the same cultures used in this footage.
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So, yeah, this lab is is pretty similar to the my other lab. They're both positive pressure labs. One thing different is the HEPA filters are basically recirculating already HEPA filtered air in this room. So the air is brought in by two two foot by two foot, uh, HEPA filtration units. Those have a pre-filter, brings in air from the next room over. It actually passes through a UV light, so that kind of kills any bacteria. Then it passes through HEPA filtration. Once it comes into this room, it's actually recirculated into these HEPA filters, so the air is super clean and also creates a positive pressure in the room. So any dust or contaminants are blown outwards instead of coming into the room.
The room is mainly focused on liquid cultures and spawn production, so there's an autoclave built into the wall. So this is a horizontal autoclave that you can load from the opposite end and unload right here directly into the lab in the clean room. So this just helps with the production of spawn, maintaining cleanliness, and also, uh, yeah, just makes it a lot easier. Then we just can store like petri dishes and stuff in here. So we have some racks, shelving units. We can also store colonizing cultures, and then we just have like some nutrition nutritional medias and ingredients in here that we can use to make up certain recipes for liquid cultures, agar slants, all that stuff. So this room is great for that.
We also have a few of these smaller autoclaves. They're All Americans, and they do great for like sterilizing small amounts of cultures, uh, liquid media or agar, stuff like that. So really this room just, uh, this lab really helps take a lot of stress off my plate because I can come in here while another lab is running and just start working on cultures and, you know, really getting into the work that I like doing. So, yeah. But all in all, both labs are very, uh, very good labs at, you know, taking care of the jobs they need to do or stuff like that.
All right, I'm kind of just, uh, giving these plates just a nice douse of alcohol. Make sure that they're nice and clean. So I'll just
loosen up my lids real nice so it's a plastic cap with a injection port so it's a self-healing injection port after you inoculate or inject a needle into it and remove it the hole will close up and also has a syringe filter that allows adequate gas exchange during the colonization so just take my scalpel actually douse the handle just a little bit just take that lid off just touching the scalpel in the middle it cools it down instantly then remove the liquid culture to the magnetic stir plate we can kind of swirl give a nice mix agitate the bit of mycelium that was placed in there hoping that little fragments of mushroom mycelium break and swirl throughout the liquid increasing the growth rate of the mushroom mycelium all right so all the inoculations have been done and at this point we can set our liquid cultures up here to colonize for about a week and we'll just check on them every day mixing as needed and once full colonization occurs We can start extracting the liquid culture into syringes and storing them for future use, or we could also just put the whole jar right into the cold storage, and that'll keep the liquid culture viable for several months. I've seen them go for well over a year, just as long as they're stored correctly, so they last quite a while. And we can see some at full colonization. This was actually inoculated March 13th, and I'll just come to it and take some liquid, uh, mycelium as needed. This is actually a shiitake mushroom. This is the 3782 82, and, uh, yeah, just a really healthy culture.
One of the things that I look for in the liquid culture is the how cloudy is it, and can you see through it? So basically, I can really see through the liquid, uh, except for the mycelium, but if you kind of see like a milky cloudiness start to occur, then that's most likely a bacterial contamination. Liquid culture is very tricky if you don't have A proper sterile technique to work with it, because it's something that you won't find out if it's not, if it's contaminated or if it's unviable, you really can't find out until you inoculate it with it. So it's, it might not be something to put a lot of hopes into for like big, big production. If you're just starting out, it might be best to start with like working on agar and being able to see the mushroom mycelium grow out right on a nice little two-dimensional surface, such as a nice solidified agar.
So this is a different liquid culture. This is another piopino one, and we can see that this one's fully colonized. A lot of the times, you'll start to see a mycelial mat develop on the surface of the liquid. This can just be re-agitated. So like if I'm was, if I were to use this right now, let's see if I can get that. So I would just basically simply agitate the mycelium by stirring it on a magnetic stir plate, and I can let that happen for about 30 seconds to a minute. The longer I let it happens, the more the mycelium can be broken down, and from there, it can just be sucked right into syringes, or you can even open up the jar and pour it directly into substrate or grain. Really, you should only be using liquid culture for grain spawn inoculations, and then from there, you can go to a substrate of your choice or of the desired species.
So then this is a turkey tail liquid culture. This is actually about a week old. Turkey till is one of the fastest colonizing mushroom species that I've worked with, usually seeing it taking over liquid culture or grain spawn or substrate within a matter of five to six days. So it's a very quick developing mycelium, a little bit slow slower of a fruiter, but it's a highly renowned medicinal mushroom, so it's a great one to add to your library or just to have in your medicine.
Cabinet you.