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Killing Trichoderma with Electrolisis. A short journal plus homemade device.


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#41 Northern Haze

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Posted 30 October 2010 - 10:09 PM

Very interesting reading :amazed:

Hang in there and keep up the good work :eusa_clap

#42 Jilkman

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Posted 30 October 2010 - 10:43 PM

It is great to see you experimenting in ways to fight contamination. Keep experimenting, but stay safe as well. It would be nice to see a more resistant strain come out of the experimentation as well.

#43 Justincase

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Posted 30 October 2010 - 11:10 PM

Thanks guys, its in the pipeline for the electrolised water, it is very tricky but once cracked it will be simple, there are lots of angles to consider. voltage, electrode size, water volume, salt content, time given for electro treatment.
Once I can find a round tooit ,will be uploading a more comprehensive line of pics for putting together the isolation implement I made for dispensing electroporation on small contam patches plus some pics on how the transferred and Re-electroporated Myc is doing (which is very well). Will be putting up more pics of the contam plate which was sprinkled in cinnamon on my other thread, the emerged Myc on that plate left over from the transfer mentioned above is steadily marching across the cinnamon and no sign of the contams covered in cinnamon. Lots to do on my list, just having a little mental break at the moment but stay tuned friends.Thanks

#44 Justincase

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Posted 02 November 2010 - 10:04 AM

Three pics from different angles of the re-electroporated Dead Trich and Mushroom Mycelium jar, taken today the 2nd of November. No sign of contams, Mycelium is growing on underside of jar as well as on the condensation forming on the jar's sides . Re-Electroporated Mycelium and dead Trichoderma was transferred to this jar on the 24th October. I can positively say that the Trichoderma fungus is well and truly dead and the Mycelium has survived electroporation twice.

Electroporated Mycelium 2nd Nov top.jpg

Electroporated Mycelium 2nd Nov Side.jpg

Electroporated Mycelium 2nd Nov Bottom.jpg

Attached Thumbnails

  • Electroporated Mycelium 2nd Nov Bottom.jpg
  • Electroporated Mycelium 2nd Nov Side.jpg
  • Electroporated Mycelium 2nd Nov top.jpg


#45 Justincase

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Posted 02 November 2010 - 12:05 PM

I havent used this device yet, but it works on the same principle as vitro electroporation (which I was initially calling electrolysis, the correct term being electroporation) . It is simple to make and use. The device was made from one 20cc plastic syringe, four pieces of electrical wires from an old computer, two lead pencils,two circuitry joiners (optional) and also two spring battery terminals for connection to the current supply (optional).Photographs are explanatory. Note, if you make this device, pick lead pencils with the lowest number such as 1B or 2B .These are made of harder grafite than the higher numbered.
The outer casing is placed over small contams and slightly embedded into the tray substrate. Normal tap water or a slightly saline solution is injected through the small side port which can also be used to extract water once the electroporation is done (filtered or purified water will impede the electrical current as it contains no salts). The inner shaft containing the grafite electrodes is lowered into the outer casing where the electrodes make contact with the water at the bottom of the casing thus creating a live electrical current through the water which will breach the outer walls of the cells structure being that of its spore and filamentation. I have also made a pair of different electrodes from Grafite that are designed to be pushed a little deeper into the substrate after injecting the contamination with water to allow a good relay of electrical current between the electrodes (see pic No.8).
The use of one layer of paper toweling before placing electrodes on a contamination is a good idea to avoid the spread of spore before disturbing the surface of the contam in question. Any more layers than one may impede the penetrability of the electrical current.

Note: Trichoderma is the only contamination I have used Electroporation on, since its growth is contained to surface, this contamination is perfect for treating in this way as opposed to a tall and fast spreading contam such as Cobweb fungus. Other surface contams are open to discovery in relation to the use of Electroporation as a means to immobilize and compromise their cellular structures.

When I first started testing Grafite as a possible use as a non shedding electrode I tested it at six volts and found there to be no shedding of Grafite particulates.I surmised that it did not shed. I wrote here that this was the case. I am happy to announce that I was wrong in suggesting this since trying out the grafite with a 12 volt charge, they did shed some particles..... I was about to smash my PC and jump through the window when I realized that Grafite is non toxic and 20 score million school children happily chew thier lead pencils five days a week. The more dense the grafite used, the less shedding of particulate 1B.

This device is prototype so if you do make it use longer wires that will reach your battery easily while placing the electrodes on your sub.
A fully charged six volt torch battery should be enough voltage to use in a contaminated Mycelium specimen to kill Trichoderma in a small volume of water. I have used 12 volts as shown on this thread but I would not do this for longer than 2 mins on account of the possibility of frying the life out of the Mushroom Mycelium.

No.1 View of surface Electroporator.jpg

No.2 Separate parts of Surface electroporator plus showing how to attach electrodes to wires thr.jpg

No.3 Above view showing wiring holes..jpg

No.4 Showing Grafite electrodes and holes..jpg

No.5 View showing barrel within outer chamber with protective cap off..jpg

No.6 View of outer chamber showing hole used to add water for surface electroporation or otherwi.jpg

No.7 Showing how to electroporate tray edging contamination without device casing using wet paper toweling.jpg

No.8 electrodes used for deeper substrate electroporation after water or saline injection of con.jpg

Attached Thumbnails

  • No.8 electrodes used for deeper substrate electroporation after water or saline injection of con.jpg
  • No.6 View of outer chamber showing hole used to add water for surface electroporation or otherwi.jpg
  • No.7 Showing how to electroporate tray edging contamination without device casing using wet pape.jpg
  • No.5 View showing barrel within outer chamber with protective cap off..jpg
  • No.3 Above view showing wiring holes..jpg
  • No.4 Showing Grafite electrodes and holes..jpg
  • No.2 Separate parts of Surface electroporator plus showing how to attach electrodes to wires thr.jpg
  • No.1 View of surface Electroporator.jpg

Edited by Justincase, 02 November 2010 - 12:30 PM.


#46 Justincase

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Posted 02 November 2010 - 12:14 PM

Acquiring Grafite from lead pencils.After burning and peeling off charcoal, lightly rub with fine sandpaper to clean surface. May be cut to size with thin hacksaw blade or folded piece of rough sandpaper. Notch where wire will be twisted around electrode to hold in place. Be gentle and patient.

No.1 Even burning.jpg

No.2 Time to snuff burning and prise charcoal.jpg

No.3 Half charcoal coating removed.jpg

No.4 Grafite extracted succesfully.jpg

No.5 What happens when the heating is excessive.jpg

Attached Thumbnails

  • No.5 What happens when the heating is excessive.jpg
  • No.4 Grafite extracted succesfully.jpg
  • No.3 Half charcoal coating removed.jpg
  • No.1 Even burning.jpg
  • No.2 Time to snuff burning and prise charcoal.jpg


#47 Justincase

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Posted 02 November 2010 - 12:26 PM

On another note, making Electro Chemically Activated water is fraught with its own problems so that is on the backburner. This has been fun and a little you know.
I have a feeling that using Cinnamon will make this thread obsolete.But it is always something to fall back on, even if the device I made proved not to be effective on substrates, one can use electroporation to revive a Trich ridden piece of Mycelium back to life in order to save a strain and try again.
I would like to test the theory of whether the Mycelium specimen I worked with now has antibodies resistant to Trichoderma or whether the exposure to Mushroom Mycelium of electrical currents does in fact strangthen cell walls of Mushroom Mycelium against Trichoderma. Perhaps I will expose the cured Mushroom Mycelium to Trichoderma spore and put my findings up here. If it doesnt work I can always treat it again.Thanks for reading. Bump it if you like it. Peace

#48 Justincase

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Posted 05 November 2010 - 07:25 AM

Pic taken today of the Electroporated Myc/Trich. Trich still dead, Myc is now growing through the middle of the dead Trich cells.Thanks

Attached Thumbnails

  • Electroporated Trich and Mushroom Mycelium specimen current growth.jpg


#49 Ethical

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Posted 05 November 2010 - 09:56 AM

Very cool, it eliminated all those other gross contams as well! Could this be applied to LC's? Agar sterilisation etc? Cutting out the pressure cooker from the equasion sounds like pure genious to me, much cheaper and more simple really. Though please oh please be careful with your health and the health of anyone else who may visit your home when growing contaminates, the endospores generated from some could prove fatal (to humans!). I'm sure noone meant to disrespect you or your experiment, but from my labtech training, any contaminates incubated on agar were handled as hazardous material. Stay safe and grow out those shrooms!

#50 iamsmiley

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Posted 05 November 2010 - 10:58 AM

i'm enjoying your creativeness in getting rid of contams,i really believe mycology as a whole is missing something big in the treatment of contams,especially trich.its nice to see someone exploring where few fear to tread.your playing with fire and some deadly things can grow on corn and peanuts as well as agar.

these are hazardous materials your playing with,that's fine so long as your taking the right safety precautions. for starters this type of research should NOT be done in your house,an outbuilding like a garden shed should be fine.wear a respirator with the right fungi rated filters sold at any safety supply store, good as long as you don't have long facial hair.don't wear your clothes in the house after working on your experiments,bag them and bleach immediately.also have a shower/bath after.a full face respirator would be best but i know they aren't cheap,getting bad spores in your eyes could cause problems but you could get away with swim goggles.there are labs out there that play with some of the worst diseases known to man but they have many layers of protection.take your safety seriously and no one will be "negative",only glad to hear your research because you'll be alive and well to tell us about this.if your unable to afford these basic precautions and don't have the appropriate outbuilding than your research may have to wait....

#51 Justincase

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Posted 05 November 2010 - 11:09 AM

Very cool, it eliminated all those other gross contams as well! Could this be applied to LC's? Agar sterilisation etc? Cutting out the pressure cooker from the equasion sounds like pure genious to me, much cheaper and more simple really. Though please oh please be careful with your health and the health of anyone else who may visit your home when growing contaminates, the endospores generated from some could prove fatal (to humans!). I'm sure noone meant to disrespect you or your experiment, but from my labtech training, any contaminates incubated on agar were handled as hazardous material. Stay safe and grow out those shrooms!


Thanks Ethical, yep I copped my Neg ratings for that outburst sweet, was a bit shell shocked from all the intense prep, plus I kind wanted this one to go into the vaults if possible, so neg comments played on my nerves. Its water under the bridge and I have thanked those who had a concern for my health, I'm fine, I have found worse growing at the back of my fridge and my brother fed me spoonfuls of dirt (which he said was Milo) under our house when I was a kid. I have a pretty strong immune sysytem, but yea, dont try this at home kids. I only kept it around to prove a point because this is important in my view.

Yes, I suppose if the endospores of the other contams got into the initial treated Mycelium then the electroporation did away with them also. I have openned that jar just about the same amount of times and in the same environment as before and havent got any contams yet, perhaps the Electro treatment somehow strengthened the cellular structure of the Myc making it harder for the spore to penetrate, but that doesnt really explain why airborne spore hasnt grown on the Maize, perhaps I have been lucky.

Yep, I have just made up a three jars of Wood shavings/Maize flour/Cinnamon juice mixed with two spoons honey and one tablespoon bee pollen/Verm. Two jars I plan to PC and the half size canning jar I am going to innoculate without PC. They have a one centimetre layer of verm that is mixed with a spoon of cinnamon powder as barrier, part of the experiment for my Cinnamon thread. I intend using this Myc to innoculate.I an thinking of exposing those to an electrical current before shooting them in order to create branching Hyphea as a possible outcome which may boost growth rate at least in the beginning stages. ( See PDF "Applied Electrical Fields Polarise the Growth of Mycelial Fungi") I believe in the experiment carried out in the above mentioned PDF, did not use an in vitro environment to expose different Mycelium to electrical feilds, hence the different outcome from my own in which imperfect fungal spore was unable to cope with the current as the Cubensis Mycelium was. Although I have tried the use of electrical current in vitro on petri plate Cobweb and it survived. It is something to do with filamentation and its possible ability to become a conduit perhaps. I am no molecular biologist...Or otherwise we are looking at singular cell versus multicellular ( and the electrically charged oxidating particles produces by passing a current through water could very well come into this) It could have been hydrogen molecules that killed them besides electroporation. This information belongs to Mycotopia now so play with it if that suits, improve upon it. Perfect it . Use it to save that strain that may otherwise go in the trash. Will have to update my grow here when there is an outcome. Gold Tops from the Western of OZ

#52 Justincase

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Posted 05 November 2010 - 11:17 AM

i'm enjoying your creativeness in getting rid of contams,i really believe mycology as a whole is missing something big in the treatment of contams,especially trich.its nice to see someone exploring where few fear to tread.your playing with fire and some deadly things can grow on corn and peanuts as well as agar.

these are hazardous materials your playing with,that's fine so long as your taking the right safety precautions. for starters this type of research should NOT be done in your house,an outbuilding like a garden shed should be fine.wear a respirator with the right fungi rated filters sold at any safety supply store, good as long as you don't have long facial hair.don't wear your clothes in the house after working on your experiments,bag them and bleach immediately.also have a shower/bath after.a full face respirator would be best but i know they aren't cheap,getting bad spores in your eyes could cause problems but you could get away with swim goggles.there are labs out there that play with some of the worst diseases known to man but they have many layers of protection.take your safety seriously and no one will be "negative",only glad to hear your research because you'll be alive and well to tell us about this.if your unable to afford these basic precautions and don't have the appropriate outbuilding than your research may have to wait....


Research is finished Smiley, thanks for your interest and concern. Much appreciated, it wouldnt have got that bad if I was believed in the first place but it was important to get the message across. I dont think I'm going to need this stuff with using cinnamon juice as sub additive and verm/cinnamon powder barrier as casing and jars tops, but that is an experimental yet to be finished. Many thanks.

#53 tricktek

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Posted 05 November 2010 - 01:37 PM

Your electrolosis idea reminds me of the Rife machine, or Rife technology. Rife invented the first microscope that allowed the viewing of live bacteria, and then subsequently learned that different electrical charges quickly killed viral, bacterial, and fungal infections, each different infection being suseptible to different electrical frequencies. Modern Rife machines can easily disinfect an entire grow room of multiple infections without harm to humans or crop.

#54 Justincase

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Posted 05 November 2010 - 09:51 PM

Your electrolosis idea reminds me of the Rife machine, or Rife technology. Rife invented the first microscope that allowed the viewing of live bacteria, and then subsequently learned that different electrical charges quickly killed viral, bacterial, and fungal infections, each different infection being suseptible to different electrical frequencies. Modern Rife machines can easily disinfect an entire grow room of multiple infections without harm to humans or crop.


Thats Cool! I have since learned that its called Electroporation.

#55 Justincase

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Posted 07 November 2010 - 07:29 AM

Something I am looking at.The Middle two are able to be read with wordpad, Thanks.

Attached Files


Edited by Justincase, 07 November 2010 - 07:31 AM.
Suggestion.


#56 iamsmiley

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Posted 07 November 2010 - 08:47 AM

Research is finished Smiley, thanks for your interest and concern. Much appreciated, it wouldnt have got that bad if I was believed in the first place but it was important to get the message across. I dont think I'm going to need this stuff with using cinnamon juice as sub additive and verm/cinnamon powder barrier as casing and jars tops, but that is an experimental yet to be finished. Many thanks.

never let your ego over ride your good judgement.did you know that Koch,one of the founding fathers of microbiology used to play around with anthrax in his house!!!

#57 Justincase

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Posted 07 November 2010 - 09:24 AM

never let your ego over ride your good judgement.did you know that Koch,one of the founding fathers of microbiology used to play around with anthrax in his house!!!


Thanks Smiley. Not sure if your message is for me. I can see how it might be but also for those who didnt read my thread properly before making negative remarks refuting my outcomes. Thanks again, I dont want to talk about it anymore, its been proved, the contams are gone, I'm fine. Koch was nuts.
The jar of treated Myc has been divided and used to innoculate a sub, will be posting the outcome.Thanks

#58 bendychicken

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Posted 07 November 2010 - 11:02 AM

Wont the mycotoxins from the trich and other badies be absorbed by the friendly mycelium?

Even if one could selectively kill off everything but the friendly stuff, wouldn't the friendly stuff still be contaminated chemically?

Not claiming to know anything, just thinking out loud......

#59 shroom_seeker

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Posted 07 November 2010 - 12:13 PM

Thanks Smiley. Not sure if your message is for me. I can see how it might be but also for those who didnt read my thread properly before making negative remarks refuting my outcomes. Thanks again, I dont want to talk about it anymore, its been proved, the contams are gone, I'm fine. Koch was nuts.

I'm pretty sure the message was for you. You are doing some interesting work, which does have potential to benefit the community. I think it's a bit premature to declare anything proven at this point, but i give you credit for what you've done and look forward to the results from your inoculated sub. You should develop a thicker skin for handling feedback and criticisms from others. The scientific method is all about critique and proving things wrong. You have lashed out in response to members who are interested in your work, and also concerned for your safety along the way. I hope you keep experimenting and sharing your work. Here's to progress, as difficult as it is to come by. :eusa_clap

#60 Justincase

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Posted 07 November 2010 - 01:37 PM

I'm pretty sure the message was for you. You are doing some interesting work, which does have potential to benefit the community. I think it's a bit premature to declare anything proven at this point, but i give you credit for what you've done and look forward to the results from your inoculated sub. You should develop a thicker skin for handling feedback and criticisms from others. The scientific method is all about critique and proving things wrong. You have lashed out in response to members who are interested in your work, and also concerned for your safety along the way. I hope you keep experimenting and sharing your work. Here's to progress, as difficult as it is to come by. :eusa_clap



Please see post #47 for proof. I have a thick enough skin as you can see from my Cinnamon thread if you care to take a look although the taunt is edited out, the patronizing comments are still there. I just find it hard to respect a comment that is unjustified and based on glazing over the facts without taking note of what is put forward. I have not lashed out at anyone because of a concern for my safety, that is appreciated. The proof is not in growing mushrooms from a sub innoculated with the treated specimen, that is just taking it a step further. The proof is that the re-treated Myc was kept on a new grow medium and grew no new contamination.One could understand my frustration at having to grow contams in order to show that if the treated Mycelium specimen was indeed still riddled with Trichoderma, surely the original treated Trichoderma would have shown signs of life well before the new airborne contams took hold. I did not want these contams in my house but since I put so much work into this and can see its worth I did what I had to do even if I put my health at risk to prove a point that I should not have had to and which I found utterly frustrating because logical thinking and careful observance of information put forward could have avoided firstly me keeping contams in my home and secondly me losing it by way of frustrating repetitive explanations.This is important, I'm not about to let an uneducated comment ruin it for other people. I would prefer it if people are going to comment, to read all info and view all pics ( taking into account the dates given for the pics and especially the time contams take to grow into consideration) before posting. This thread is not "all about me" in the least. Its about showing something that can be done about surface contams or to save a strain if that is all one has left to work with.
Just to note again, the sub I have innoculated....if it somehow gets a contam in another area of the sub, then this whole deal is bunk sorry but if one cant make a logical conclusion by taking all facts into play and viewing
the pic and date for post #47 then I give up, yep my skin is only so thick and after a while people get worn down. And this is not me lashing out but I'm kinda sick of talking about this and the whole "I put my life at risk" thing, points have been made, I dont understand why people are repeating themselves, its old hat..that stuff was said in the first page, I'm not stupid.
I feel like when I was a kid and my parents would come in and start shoutin make yer bed! Make yer bed! Id still be asleep! Another thing is...I'm not telling anyone they have to do this, this is what I'm doing I just thought I would share is all. This site is supposed to be for adults, I dont see kids reading it and going "Oh good idea, think I'm gonna grow a whole bunch of contams like this dude :amazed:" I suppose the most conclusive evidence it works is when I fruit the Mycelium but for me it is already proven (twice).I just want to move on now and finish my grow with as little fanfare as possible which is what I most desired for this thread in the first place.

Thanks for the credit you've given, your positive criticism as well as your encouragement and balance of judgment.Peace




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