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


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#1 Justincase

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Posted 23 October 2010 - 12:57 PM

This thread is to show how Trichoderma can be killed using Electrical voltage in water. I will try to keep this as simple as possible and also have built a small device that is easy to make that you can use to spot kill Trichoderma and possibly other contam outbreaks on substrate.
Firstly I would like to share a document on treatment of bacteria in an aqueous solution using electrolisis.
http://www.ecaa.ntu....suspensions.pdf
Here are pictures from my experiment where I took a piece of Mushroom Mycelium which was mostly overtaken by Trichoderma fungus and exposed it to ten minutes of Twelve volts of electricity in shallow water.As you can see, the Trichoderma has died as well as its spores and the Mushroom mycelium has survived and re-emerged. The Piece of treated Mycelium was placed on a plate of Maize flour paste it has been five days now and the plate has been exposed to air a number of times and although the Maize paste has accumulated some new occurrences of Trichoderma and red mold, these contaminations have not come from the killed off piece of Trichoderma infested Mushroom Mycelium but from air exposure of the Maize paste.
No.1
1 Trichoderma contaminated Mycelium directly after ten minutes of 12 volt treatment in H2o.jpg

No.2
2 Directly after shock treatment the specimen has been embeded in Maize flour.jpg

No.3
3 One day after placing in Maize flour specimen has sunken into substrate due to excess moisture.jpg

No.4
4 Three days in Maize flour.jpg

No.5
5 Mycelium growing out from under Dead Trichoderma 22 Oct 4 days since embeding in Maize flour.jpg

No.6
6 Showing that the green coloring is not caused by electrolisis of the terminals.jpg

No.7
7 Five days Healthy Mycelium growing from under dead Trich as well as new airborne contaminants .jpg

Attached Thumbnails

  • 7 Five days Healthy Mycelium growing from under dead Trich as well as new airborne contaminants .jpg
  • 6 Showing that the green coloring is not caused by electrolisis of the terminals.jpg
  • 4 Three days in Maize flour.jpg
  • 5 Mycelium growing out from under Dead Trichoderma 22 Oct 4 days since embeding in Maize flour.jpg
  • 3 One day after placing in Maize flour specimen has sunken into substrate due to excess moisture.jpg
  • 2 Directly after shock treatment the specimen has been embeded in Maize flour.jpg
  • 1 Trichoderma contaminated Mycelium directly after ten minutes of 12 volt treatment in H2o.jpg

Edited by Justincase, 23 October 2010 - 01:35 PM.


#2 Justincase

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Posted 23 October 2010 - 01:04 PM

I have made an isolated electrolisis device made from household objects which can be used for spot killing contams on tray substrates especially during pinning stage and will be showing you how to make one and also how to use it.

1 View of contamination electrolisis device.jpg

2 Side view of Contaminant electrolisis device.jpg

Attached Thumbnails

  • 1 View of contamination electrolisis device.jpg
  • 2 Side view of Contaminant electrolisis device.jpg

Edited by Justincase, 23 October 2010 - 01:21 PM.

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#3 hyphaenation

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Posted 23 October 2010 - 01:21 PM

The agar plate you show with myc coming from under the trich ... that looks to me to be trichoderma mycelium. I'm sure your well aware that trichoderma mycelium is also brilliant white , before it sporelates , which is the green we see.

Could be Cubensis mycelium , but looks a lot like trichoderma mycelium. I'm certainly not trying to discourage you.

Interesting experiments ! Keep it up.

#4 Justincase

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Posted 23 October 2010 - 01:32 PM

The agar plate you show with myc coming from under the trich ... that looks to me to be trichoderma mycelium. I'm sure your well aware that trichoderma mycelium is also brilliant white , before it sporelates , which is the green we see.

Could be Cubensis mycelium , but looks a lot like trichoderma mycelium.

Interesting experiments ! Keep it up.


Thankyou My camera is fucked, so you wont see the fluffy mycelium I suppose.How about some math, Myceliums been growing for four days, is it green? No.How long does Trich take to spore?

Edited by Justincase, 23 October 2010 - 01:58 PM.


#5 Freaky

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Posted 23 October 2010 - 02:00 PM

:cool:

You may want to make us some fresh, sterile agar plates. The maize flour paste might be allowing other contaminates to skew the results.

I do think you had healthy mycelium starting out, but all the other contaminates growing make it hard to tell.

If possible, give it another go and try some transfers right when you see the first signs of new mycelium growth from under or around the trich pieces.

#6 Justincase

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Posted 23 October 2010 - 08:42 PM

:cool:

You may want to make us some fresh, sterile agar plates. The maize flour paste might be allowing other contaminates to skew the results.

I do think you had healthy mycelium starting out, but all the other contaminates growing make it hard to tell.

If possible, give it another go and try some transfers right when you see the first signs of new mycelium growth from under or around the trich pieces.


Thanks Freaky, I think the plate including contams gives a good representation of what Trich looks like as well as Mycelium of Mushroom side by side. If you really study the pics you can see that the Mycelium growing in the middle was there well before the outer spots of Trich showed up.Those Trich spots have turned green and the centre Mycelium is still white and fluffy.If the Mycelium in the centre of the plate was Trich it should be green by now. Is this not a
logical conclusion? For the Centre cloud of Mycelium was there before the outer contams of Trich showed up.I am going to use this plate and its contams to show how to use the isolating electrolysis instrument I have made.

Edited by Justincase, 23 October 2010 - 09:54 PM.


#7 Justincase

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Posted 24 October 2010 - 03:37 AM

Hello, this is a picture of what the plate looks like today, I will move the Mycelium to a new plate of Maize paste substrate and allow it to grow. I am wondering if it will now have antibodies resistant to Trich. As you can see in the pic, the Trich and red mold is quite distinctive as opposed to the Mycelium which is still white and fluffy and advancing across the surface of the plate, it is six days now since it first re-emerged from the half contaminated specimen after electrolysis.I can understand that people may be thinking "Yes but that trich spore must have come from the Trich you just gave electrolysis to so how can you say you killed it?" This is not true, the spore came from the air as the plate has been exposed many times, for starters I left the plate with no lid underneath a lamp to dry off excess moisture from the Maize paste I made up, there has been ample opportunity for airborn contams to get onto the plate.

No.8 Six days after electrolysis of Trichoderma contaminated Mycelium.

Attached Thumbnails

  • 8 six days after electrolysis.jpg

Edited by Justincase, 24 October 2010 - 03:44 AM.


#8 director of sound

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Posted 24 October 2010 - 05:07 AM

ive found success with UV and H2O2/salt. is sprinkle salt on the infected area and put dome 10% H2O2 on the spot then put in a homade steralizer with a 20W UV lamp in it for 12hr. the myc survives but any surface contam is toast.
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#9 Justincase

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Posted 24 October 2010 - 07:31 AM

ive found success with UV and H2O2/salt. is sprinkle salt on the infected area and put dome 10% H2O2 on the spot then put in a homade steralizer with a 20W UV lamp in it for 12hr. the myc survives but any surface contam is toast.


Cool! Im going with this instead, its going to save me allot of bother.Thanks!
My idea still would have run into problems on tray edging. BTW what is a homemade sterilizer and how many times do you have to do this from say pinning stage to end result if the Trich shows up around pinning time or one week before? Electrolysis is good for a last resort to save a specimen of mycelium for rejuvenation if it is all you have left after a jar has been contaminated but I cant be bothered putting the energy into convincing anyone it works anymore. I think it would be helpful if you were a bit more specific such as saying how close you place the light, whether it needs to be a specialised UV lamp or just globe in an ordinary lamp, does the heat from the lamp dry out the sub. Maybe you could do a thread about it or leave a link here if there is one explaining it further. Many thanks Director Of Sound
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#10 Freaky

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Posted 24 October 2010 - 11:12 AM

Good info, and yes, after 6 days it should have returned to green around the original piece.

Let us know if you have any success with the UV sterilization and post your build and experiments :)
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#11 Freaky

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Posted 24 October 2010 - 11:12 AM

Good info, and yes, after 6 days it should have returned to green around the original piece.

Let us know if you have any success with the UV sterilization and post your build and experiments :)

#12 loudenbomber

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Posted 24 October 2010 - 12:11 PM

Good post, lots of useful information and ideas .. +rep ,.



I believe your mycelium had returned. I'm curious as to weather an isolate of that myc would show more resistance to trich contam. This possibly could be something that wouldn't be that difficult to do to build up a nice resitant isolate to use.


Also , I'd also very much like to see DOS's build on UV steralizer, that would be perfect for aluminum plate subs when possible contams get in around the edges.
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#13 TVCasualty

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Posted 24 October 2010 - 12:13 PM

No.8 Six days after electrolysis of Trichoderma contaminated Mycelium.


I would be very careful about letting petri dishes contaminate to such a degree in my house. Actually, I wouldn't do it in my house at all.

Looks like you got some aspergillus (maybe), some penicillium, and maybe lipstick mold (aka Sporendonema purpurescens or Geotrichum candidium) on that one dish.

In concentrations large enough to see on a petri dish these contaminants can become a very serious health hazard. Aspergillus niger is a common cause of fungal ear infections and some penicillium species can produce potent mycotoxins.
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#14 Dr.Hallucination

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Posted 24 October 2010 - 12:34 PM

I agree with TV,that is the nastiest dish I have seen in a long time,be careful with that sucker...

You should have tossed that dish out a long time ago,or as TV said,not have started it in your home in the first place.But its your home,your health,and all future projects at stake,not mine so have it er...


The only thing I have seen so far in this thread is your machine is not doing a thing to help that dishes cause in anyway.

I am not saying the project or idea is dead,just that damn dish is.Toss it out for for the love of god.Start the project over,you have all the time in the world.Just not with that dish.

#15 Justincase

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Posted 24 October 2010 - 01:22 PM

I agree with TV,that is the nastiest dish I have seen in a long time,be careful with that sucker...

You should have tossed that dish out a long time ago,or as TV said,not have started it in your home in the first place.But its your home,your health,and all future projects at stake,not mine so have it er...


The only thing I have seen so far in this thread is your machine is not doing a thing to help that dishes cause in anyway.

I am not saying the project or idea is dead,just that damn dish is.Toss it out for for the love of god.Start the project over,you have all the time in the world.Just not with that dish.


Yea its pretty gnarly, But since I was having trouble getting the message across I let it go extra time. Please for the love of weed analyse the facts before you tell me my electrolisis is funk. How many times do I have to say before I bang my head against the wall, okay in capitols now for the blind.

THE PIECE OF TRICH INFESTED MYCELIUM WAS TREATED TO A 12 VOLT ELECTRICAL CHARGE IN WATER THEN MOVED TO A PLATE OF MIAZE MIXED WITH WATER. THE MIAZE PLATE WAS OBSERVED AS THE MYC CAME BACK TO LIFE AND THE TRICH STAYED DEAD THE NEW CONTAMS DIDNT COME FROM THE DEAD TRICH BUT FROM OPEN AIR EXPOSURE OF THE PLATE.THE CONTAMS HAVE COME AND GONE TO SPORE , THE MYC IS STILL THERE AS YOU CAN SEE AND AS FREAKY ALSO HAS AGREED THE MYC HASNT TURNED GREEN AND AT THE RISK TO MY OWN FUNKING HEALTH TO MAKE A POINT WHICH I SHOULDNT HAVE TO I WILL UPLOAD A SEVENTH DAY PIC TO PROVE WHAT I ALREADY HAVE THAT IT IS MYC ONCE AND FOR ALL AND IF THE TRICH WAS STILL ALIVE ON THE SPECIMEN I ELECTROCUTED, IT WOULD BE GREEN OR WHITE BUT NO, ITS CELLS GOT FUNKED OVER IN THE ELECTROLYSIS I KICKED ITS ASS WITH. ALSO TWO WIRES ARE NOT A MACHINE AND NEITHER IS THE OBJECT I AM HOLDING UP IN THE PIC, ALL IT IS IS A SYRINGE WITH THE END CUT OFF, TWO WIRES DOWN EACH SIDE, YOU PUT THE END OVER TRICH ON A TRAY SUB,PUT SOME WATER IN, LOWER THE CENTRE IN SO THE WIRES ARE CONTACTING THE WATER AND HAVE THE OTHER END OF THE WIRES HOOKED UP TO A BATTERY "NOT A MACHINE" MACHINES HAVE MOVING PARTS! FUNKS SAKE IF ANYONE TOOK MY WORD FOR IT i WOULDNT FUNKING HAVE THIS STUPID DISH IN MY FUNKING ROOM MAKING ME SICK FOR FUNKS SAKE!! why are people so damn negative and always gearing thier minds for negative outcomes, at least a few sane minded people here have had the damn brains to see that it worked . I salute you!

Edited by Justincase, 24 October 2010 - 01:36 PM.


#16 Justincase

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Posted 24 October 2010 - 01:30 PM

Good post, lots of useful information and ideas .. +rep ,.



I believe your mycelium had returned. I'm curious as to weather an isolate of that myc would show more resistance to trich contam. This possibly could be something that wouldn't be that difficult to do to build up a nice resitant isolate to use.


Also , I'd also very much like to see DOS's build on UV steralizer, that would be perfect for aluminum plate subs when possible contams get in around the edges.


Thanks Loudenbomber I appreciate it. And yes it very well may do so, I am taking the tray out tonight , will photograph it one more time tommorrow and remove the mycelium before destroying the rest with bleach , I may even leave it in there (outside)to see if it is impervious to Trich, if it isnt I could always give it volts again. I was going to show how to make the thing im holding up but no one seems to be getting it. Whole thing is doing my head in.:gah:

#17 Justincase

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Posted 24 October 2010 - 01:49 PM

Dr hullucination I apologise but do the math, if it were Trich it would have turned green but now and it isnt Cobweb I can assure you. The other trich and red mould you see are new contams from airbornes, nothing to do with the dead trich and no I havent used electrolisis on those, I left them there to show the difference becuase people were having trouble realising the white surrounding the dead trich was indeed Mycelium .

#18 Justincase

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Posted 24 October 2010 - 02:01 PM

Good info, and yes, after 6 days it should have returned to green around the original piece.

Let us know if you have any success with the UV sterilization and post your build and experiments :)


Thanks Freaky, dont have the cash to run out and get a UV lamp at the moment. Am happy myself with results and will upload closer pics explaining the device I made and its proper use. Am taking a break from all this for a bit.

#19 Freaky

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Posted 24 October 2010 - 02:34 PM

TV and DrH make very important points about your health and safety with experiments and contaminations being present. Please do take their words given out of respect to your well being.

I'm always happy with members willing to post experiments and findings whether tey turn out good or bad.

#20 Justincase

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Posted 24 October 2010 - 10:15 PM

TV and DrH make very important points about your health and safety with experiments and contaminations being present. Please do take their words given out of respect to your well being.

I'm always happy with members willing to post experiments and findings whether tey turn out good or bad.


Yes I agree with concern for my health but I feel disrespected because I can tell they arent taking all of the facts put forward including pics and analysing them properly before making an educated post on my thread which ends up downing the whole outcome and results thus making it seem like a flop when the truth is there for anyone with some wit to figure it out. I dont have a problem with what TV said( although that dish has the lipstick mould, Trich and only Mushroom Mycelium )but when I put time and effort into this and someone flips a half assed comment after glazing over it( and I can tell thats what was done because I havent even used the implement I made yet but it follows the same theory as in my first experiment that passing an electrical current through Trich erupts is cells) it really fires me up. The only reason I have this contam hanging around was because I was forced to prove the point further about the time Trich takes to spore as opposed to the Myc which was there before the Trich and is still as fluffy and white as a fairies ass. I get the feeling because Im a noob no one is going to take this seriously. Kinda like a decent artist never gets recognition because they never went to univeristy and got a degree in how to paint a cowshit blue and nail it to a board. I feel like Im being called a liar, but what would the point be of lying, its not like I would get anything out of doing that. I guess it comes down to I know it works and I was willing to share , if someone needs convincing further, too bad. I dedicate this thread with those born with a brain in all due respect. Im not here to dance on peoples threads looking smart and smug, Im here to share and learn.




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