Killing Trichoderma with Electrolisis. A short journal plus homemade device.
Posted 05 October 2017 - 08:56 PM
I read your first post at 4:30 am after a very long day. I apologise.
Yes Sandman you are right absolutely that sterile technique is the key to non contamination. Correct x3.
I'm doing this for a few reasons. One of those is because "I love it"
Yes we try to be as sterile as possible. But sometimes contaminants get into projects.
This won't only kill trichoderma in an LC but most other if not all contaminants.
I believe it will also cause faster growth in mycelium.
This is just another tool.
I'm happy to test it myself and already have with very positive results.
Nobody has to use it. This is just what I'm doing for my own interest and sharing my findings. I have been contacted by an account holder here to say he tried it and it worked.
I would use this technique as a last resort to saving a culture/species when all other options are lost.
Say you make an LC from a print you thought was sterile. You have inoculated some jars but they all contam.
You either make agar plates and try to outrun the contams. A long process.
Or take some of your LC and treat it in this device which knocks out your contains and leaves you with clean LC which you can then plate test.
I don't see how this tool could be a waste of time.
I see it as a viable way of ensuring the sterility of tissue culture also as well as a growth booster for the tissue sample before being placed on agar or into liquid culture.
The design of my treatment apparatus is functional. But I have a more simple design that will be easier to extract from. I'll make that soon.
Again. I apologise for taking your comment negatively. I'm very stubborn. I don't have time for doubt once I've made up my mind on something.
Posted 05 October 2017 - 09:08 PM
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Posted 05 October 2017 - 09:10 PM
With all respect Sandman. I'd be delighted to hear how you know that my device (no moving parts therefore not a contraption) won't kill contaminants.
Have you studied irreversible electroporation?
Effect of charged particles on fungal spore and bacterium in aqueous solutions?
I've read papers on the technique applied at 1.5 volts which were proof of a destructive interaction between electricity and mould spore/bacteria.
I am using 12 volts and have already shown that mushroom mycelium recovers from this exposure.
Posted 05 October 2017 - 09:18 PM
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Posted 05 October 2017 - 09:27 PM
Posted 07 October 2017 - 12:24 PM
I've loaded a mix of half wood chip tea and spores obtained by shaking a much handled dry
P. Subaeruginosa into a specimen container of water. Close to 1.5 mls of each.
I'll allow the spores to germinate and grow liquid culture inside this before introducing a twelve volt current through it. I can say without a doubt that this spore solution is contaminated but I'll happily introduce some of the solution to agar before I begin the irreversible electroporation process on the Contaminants within the LC.
I like this design a lot more than the last as it focuses the current inside a tighter space. The free radical electrons will hit their target more efficiently. Plus you can put it together in five minutes from a clear generic pen barrel.
I would have liked to have the graphite electrode fitting the opposite end but I needed one breather hole for gases and one for the copper wire negative electrode. I melted those into the pen barrel stopper with a hot tack.
The plastic coated wire is just for hooking the vessel to stand it up straight while filling and using my camera and of course for pulling out the stopper easily.
Just keeping you up to date. (Silent watchers included).
Note the colors of wires are backwards. Red is usually for positive wires. This is just a convenience for me (mainly because of the length of wires and I was being lazy) . The electrons actually flow from the negative electrode and ions flow from the positive if I am correct.
I am going to go through all of my saved information and take notes so I can clearly convey what exactly is happening when current passes through this.
BTW since this is such a small area to cover I'm not using an electrolyte. There should be enough molecular energy in the wood tea to allow current transfer through the water and as the electrodes shed ions/electrons the conductivity of the solution will build.
Reading so much. Very fascinating what this does. I think I have enough info accumulated to be able to explain so I'll being formatting a concise explanation.
I just have to wait for the LC to grow then I'll hit it with the 12 volts.
Btw the pic of it wired to my battery shows the copper wire out of the water. It would be in be at least one cm if I were completing the circuit and electrolysing the solution of spores and wood tea.
Its just sitting upright in a box with foil over the holes for now.
Treatment will be done in a glove box.
I'll have go at making a separate thread for this when I've proved it enough times. This thread has the wrong heading. More of a testing thread really..Thanks for your patience. Questions. Go ahead and ask.
Edited by Justintime, 07 October 2017 - 12:41 PM.
Posted 07 October 2017 - 12:52 PM
How current flows
Copper is a good conductor because, like other metals, it contains free electrons. Free electrons are also known as conduction electrons. Each copper atom provides a single free electron, so there are as many free electrons as atoms.
Free electron concentration in copper n = 8.5 × 1028 per m3
When a voltage is connected across a piece of copper, it pushes the free electrons so that they flow through the metal – that's an electric current.
Interactive graphic of electrical conduction
Interactive graphic of electrical conduction
Picture 3.4 Close the switch to apply voltage: electrons start to flow through. Increase the voltage and they drift faster.
Notice that the electrons start to flow as soon as the switch is closed. The message to get them moving is instantaneous (in fact it travels close to the speed of light). However, the electrons themselves travel much more slowly. So how does the current come on everywhere as soon as the switch is closed?
It is because the free electrons are already spread through the wire. As soon as the switch is closed, there is a force on all the electrons, which gets them moving. It's a bit like a bicycle chain. As soon as you start pedalling, the back wheel starts to turn. The force on the back wheel is instantaneous even though the individual links are travelling at a visible speed. But because the links are already spread around the chain 'circuit' they all start to move at the same time.
The animation above is slightly deceptive because it shows the electrons as being stationary when there is no voltage.
In reality, even when no current flows through a piece of copper, the free electrons are moving rapidly about. Their speed is about 106 m s-1; that's 3000 times the speed of sound in air! However, since they are moving at random, there is no net flow of electrons in any particular direction and so there is no current.
When a voltage is applied, the electrons gain an additional velocity, so that there is a net flow along the wire. This extra velocity is called their drift velocity. Here's a way to picture this:
Think of a swarm of bees. They are all milling around the hive. Each bee is moving, but the swarm stays still. Now one sets off and the rest follow. The swarm is still a milling mass, but overall it moves away from the hive. The free electrons in a metal are like the bees; it takes a voltage to make the mass of electrons move through the wire.
Graphic of current
Picture 2.5 Conventional current flows in the opposite direction to the electrons.
Electrons are negatively charged. They flow from negative to positive in a circuit.
Conventional current flows the other way - from positive to negative
Posted 12 October 2017 - 12:08 PM
Hehe Okay this is my last post on this thread. I showed this works a few pages in so I've been feeling like I'm just flogging a proverbial.
But I'll leave a big fat juicy clue for anyone serious about understanding this process. And that is this sentence.
"Ions scavenge electrons from lipid structures causing chain reactive oxidation"
Posted 15 October 2017 - 08:59 AM
Edited by Justintime, 15 October 2017 - 09:01 AM.
Posted 15 October 2017 - 09:07 AM
Not only does electrolysis create oxidising ionic reactions which destroy phospholipid structures such as cells .
Electrical current in aqueous solution causes irreversible electroporation. This is when the hydrophobic phospholipid membrane surrounding the cell rearranges itself creating a pore or hole in the cell wall allowing the liquid to penetrate the cell wall. In instances of prolonged exposure to electrical current this hole stays open (irreversible electroporation) The cell is flooded and dies. Especially if the electrolyte used I'd antifungal/antibacterial.
That should do it. Now I will be immortalised with the mushroom Gods and all will silently chant my name in remembrance of the greats. Twooit deeedle peeedle pompom zip meadle!
Edited by Justintime, 15 October 2017 - 11:19 AM.
Posted 15 October 2017 - 10:11 AM
This is why I am allowing the mushroom spores to grow mycelium before treatment via electrolysis/irreversible electroporation/ ionic oxidation. There is a strong possibility that the treatment will destroy mushroom spores.
Just revising. The treatment is not damaging to mushroom mycelium due to the presence of antioxidants which willingly give over electrons to ions thus avoid ing oxidating chain reactions of cell walls.
Electrical currents polarise mycelium.
The Lipids of Agaricus Bisporus
Edited by Justintime, 15 October 2017 - 11:18 AM.
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Posted 03 November 2017 - 08:16 AM
Simple as it gets. Graphite electrodes from a HB pencil (set on fire to burn away wood. Graphite withstands 2000 degrees).
No electrolyte used (it was found that the wood tea contained enough ions (salts) for current movement.
Bubbles forming at electrodes assumed to be hydrogen gas and oxygen (as no literature available on gases formed between two graphite electrodes in ionic solution , I cannot say for sure. Not important but as a mostly alkaline solution used it will be hydrogen and oxygen. Possibly carbon dioxide due to carbon content of electrodes. So h20 +carbon= carbon dioxide.
I see we have a member here who is trained in Chemistry. Feel free to chime in if you're here .
Receptacle is a contact lenses container (double ended) Hole melted through centre barrier.
Stopper ends taken from syringe parts. Holes for electrodes made using leather hole punch.
12volt direct current power supply.
Treatment and testing ongoing. I am happy with this design. It's nice and compact. I require a small dental cleaning kit for vibration. This device will be housed upon the sonic cleaner for this reason. To facilitate the movement of free radicals (ions).
Note: If the solution were distilled water, the current would not move between electrodes. I have tested calcium carbonate as an electrolyte and it works in this small space( source: cuttlefish bone). Non toxic,alkaline. Mushroom friendly. Not a salt.
BTW check the bubble forming at each electrode.
Will make a much neater concise thread for this once everything complete.
Anyways just letting anyone following where I'm at with this. Time to start agar tests. So many distractions lately. Peace
Posted 03 November 2017 - 10:52 AM
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