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OfflineKryptos
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Autoclaving substrate with high motor oil content (<10%)
    #23520130 - 08/07/16 07:58 PM (7 years, 5 months ago)

I'm fairly certain that this thread belong here, and not the regular cultivation forum...If I am wrong, please correct me/move this thread. I don't know if I can be considered an "expert" cultivator, but I've been selling homegrown oysters at farmers markets for a few months, and I've got oyster growing down to where I'm pretty comfortable and know how to deal with all the issues I've encountered so far. Now, I want to experiment with some mycoremediation possibilities that oysters seem to be quite comfortable with.

I want to start with used motor oil. Mostly because I have a few gatorade bottles full of the stuff that I haven't taken to the local hazwaste facility yet. I wanna make clear that any oysters grown on such contaminated substrate will not be eaten, and will likely go straight onto the compost pile. Or possibly get reprocessed into new substrate by dehydration/pulverization and re-sterilization. I haven't tried that either, but I don't see particularly why it wouldn't work.

So far, my plan is fairly simple: make a set of jars that have varying amounts of motor oil in them, from 0.1% to 10% by weight motor oil with the rest being a mix of mostly straw and a little WBS. I know that there have been tests done with pouring oil onto colonized straw logs, but I don't really have an environment conducive to such tests, mostly because I don't want standing oil in my fruiting environment/house.

The problem that arises in my mind would be the safety of autoclaving motor oil. While oil is generally heat-stable (that's one of the reasons it is used) I am not particularly interested in testing this out in my kitchen. Or the local lab. I am not too worried about possible polymerization results, according to my research, oysters should be able to eat hydrocarbon polymers. I could theoretically try to sterilize substrate ahead of time, and then add varying amounts of oil right before inoculation in a BSL cabinet.

So, three questions:
1) How smart is it to autoclave oil?
2) Is it even possible to reuse fruitbodies as new substrate? (I expect mediocre-to-severe productivity losses and a much lower BE)
3) (Assuming the answer to #2 is "yes") Should the resulting mushrooms be remashed into new substrate to further reduce possible environmental impacts, or would they be fine going into the compost pile?

The third question will likely be answered by doing some analytical testing of the resulting fruitbodies. I was just wondering if anyone knew about this already.


Edited by Kryptos (08/07/16 08:00 PM)


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Invisiblepablokabute
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: Kryptos]
    #23520236 - 08/07/16 08:26 PM (7 years, 5 months ago)

subbing. great ideas you got there going...


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Offlinedrake89
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: pablokabute]
    #23520418 - 08/07/16 09:20 PM (7 years, 5 months ago)

I think if you were gonna do this, don't reuse your pressure cooker for edibles amd do it outside.  There could still be some gasoline in your oil.


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Offlineinvitro


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Re: Autoclaving substrate with high motor oil content (<10%) [Re: drake89]
    #23520757 - 08/07/16 11:37 PM (7 years, 5 months ago)

http://voices.nationalgeographic.com/2015/08/03/heavy-metals-in-motor-oil-have-heavy-consequences/

As motor oil circulates through a car’s engine it picks up heavy metals such as arsenic, lead, cadmium, copper and zinc, as well as other toxins.

Need I say more?


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OfflineKryptos
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: invitro]
    #23521444 - 08/08/16 09:17 AM (7 years, 5 months ago)

The heavy metals thing I am not too worried about. They're called heavy for a reason (yeah, I know the reason is high atomic weight, but they're toxic because they tend to build up wherever you put them, instead of volatilizing or getting washed out). I actually specifically wanted to use *used* motor oil to test how well oysters perform as bioaccumulators of toxic metals.

Having done some more thinking, I'm leaning towards not autoclaving the oil (I'd be using a lab autoclave at the local Uni, not my PC), but just autoclaving the sub and then pouring oil into jars after in a sterile environment. I'm fairly certain motor oil would be auto-sterile, as a result of water and oxygen exclusion. I may also filter-sterilize the oil, considering the relatively small volumes involved.

Anyway, my plan right now is fairly simple: 6-7 half pint jars (or as many as I have free tonight, grain is already soaking), with a substrate that is 5:1:1 mulched straw:fresh compost:WBS (my normal spawn runs are 6:1 straw:WBS. I figure a little bit of compost for funzies, plus I haven't taken anything out of my pile in some time. These will be sterilized with regular ported tyvek filter lids. Then, I will use a BSL cabinet to pour varying amounts of motor oil into each jar, between 0.1 and 10%. I will then leave the jars alone for a week to check if oil is sterile and make sure nothing funky starts growing. Assuming nothing starts growing, I will inoculate with 1ml of heavily colonized LC (I know, excessive, but I figure the mycelium could use all the help it can get with the motor oil), and then observe colonization times. I expect some variability caused by varying LC concentration, but overall I expect the jars with more oil to colonize slower. Once the jars are colonized fully and consolidated, I will spawn them as cakes, as they already have a good ratio of bulk sub to spawn. Flushes will be collected and weighed per jar after I see the first spores drop. The reasoning here is just standardization. Normally, I collect fruitbodies when they "look ready", not any quantitative measure. First spore drop would give me some semblance of standardization and control. Cakes will be flushed until they are no longer capable of flushing, regardless of contamination. Finally, the first, middle, and last mushrooms produced will be prepared and analyzed by UHPLC/MS to test for long chain hydrocarbons and metals. Cakes will be similarly analyzed after they finish flushing.

I haven't really thought about how I'm going to do sample prep at the end yet, as I don't know any solvents that can extract and quantify heavy metals reliably without derivatization. The analysis of long-chain hydrocarbons would be simple using regular organic solvents (probably ACN, that's what I usually use in my HPLC). What do you guys think? Am I missing anything?


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InvisiblebodhisattaMDiscordReddit
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: Kryptos]
    #23521710 - 08/08/16 11:05 AM (7 years, 5 months ago)

Motor oil is definitely not sterile. It accumulated dust while it was in the environment. Relative low CFU I'm sure but I would not call it sterile or near sterile.

I would sterile filter it. Pump thru a .2 micron filter


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Edited by Trusted cuItivator (08/08/16 11:06 AM)


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Offlinehorneeowl101
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: bodhisatta]
    #23528132 - 08/10/16 12:40 PM (7 years, 5 months ago)

Excited to see where this goes


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Invisibleflyontoast
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: Kryptos]
    #23559545 - 08/20/16 10:02 AM (7 years, 5 months ago)

Quote:

Finally, the first, middle, and last mushrooms produced will be prepared and analyzed by UHPLC/MS to test for long chain hydrocarbons and metals. Cakes will be similarly analyzed after they finish flushing.

I haven't really thought about how I'm going to do sample prep at the end yet, as I don't know any solvents that can extract and quantify heavy metals reliably without derivatization. The analysis of long-chain hydrocarbons would be simple using regular organic solvents (probably ACN, that's what I usually use in my HPLC). What do you guys think? Am I missing anything?




Hope this experiment is going well so far. I am concerned for your compost pile. My understanding is that the fruit/mushroom will for sure contain heavy metals, but the substrate and mycelium won't. As you already know, most fungi are natural remediators (mycoremedeation), so the mycelium can clear large areas of contaminants and heavy metals, but everything that the fungi cannot break-down gets concentrated into the fruit. In nature this allows for plant life to return to area that the fungi has "scrubbed", sacrificing only a tiny area where the mushrooms decompose to the contamination. So, in your experiment, the substrate would theoretically be drained of any metals and, if all the oil is broken down too, you could compost the substrate. But you'd still be stuck with mushrooms full of dangerous materials that you shouldn't return to nature, and especially not your garden(?) compost. I'm curious to see the results of your tests and lemme know what you do with the mushrooms if you find a solution since I'd like to try it too if you get good results.

Here's Stamets on how this works when it comes to mycoremediation of radioactivity:
https://www.permaculture.co.uk/articles/how-mushrooms-can-clean-radioactive-contamination-8-step-plan


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:mushroom: My trade list :mushroom:
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Agaricus Bitorquis, Agaricus Augustus, Agaricus blazei/subrufescens, Stropharia Rugoso-annulata, Clitocybe Nuda (blewits), and any species or other genus that you think work outdoors.
Also, any commercially viable Pleurotus, cold or hot strains.
Thanks for the Q&A, trades, and all the posters & teachers that have come before us  :skol:


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Invisibleenlightened seed
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: Kryptos]
    #23605920 - 09/02/16 07:49 PM (7 years, 4 months ago)

if you are successful, send your specimens to a lab to have tested to see what they contain perhaps?  interesting subject though....


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OfflineKryptos
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: enlightened seed]
    #23624029 - 09/07/16 10:15 PM (7 years, 4 months ago)

I have a full analytical lab with Fischer's new flagship HPLC at my disposal. Sadly, I also have the associated grad school and the semester just started, so the project was put on hold after initial prep. Once everything settles down a bit I will hopefully continue.

Assuming I get anything interesting in preliminary results, I plan to look into the possibility of securing funding through collaboration with the local mycology prof.


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OfflineMarcTheMushroom
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: Kryptos]
    #23755705 - 10/20/16 06:28 PM (7 years, 3 months ago)

Physics and thermodynamics

Autoclaving soil with oil in it would be perfectly fine!!!


you are still just putting water in the bottom of the pot and boiling it at 15psi and it will only get upto 121'C


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InvisiblebodhisattaMDiscordReddit
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: MarcTheMushroom]
    #23756035 - 10/20/16 08:26 PM (7 years, 3 months ago)

you have to worry about the partial pressure of anything other than water if it goes into gas phase in an autoclave.


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OfflineLennybernadino
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Re: Autoclaving substrate with high motor oil content (<10%) [Re: bodhisatta]
    #23760842 - 10/22/16 12:38 PM (7 years, 3 months ago)

Well if there is any galoline or naptha still in the oil it will go into a gas state, but that is probably gone already right?


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