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Invisibleredrum
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Constant Contamination
    #22470696 - 11/03/15 11:56 AM (8 years, 2 months ago)

Hey guys,

I tried twice to cultivate following pf tek to a "T". Pink mold is a constant issue.

Washing the jars with antibacterial soap, spray with bleach, and rinse again. Prepare the substrate and in the cooker they go. 15psi for 1hr.

Pot cools down for about 6 hours. Prepare room by spraying EVERYTHING down with bleach and lysol, including pressure cooker before opening and letting it sit for 10 min. I even wore rubber gloves and sprayed them. I opened the cooker and sprayed bleach solution inside on tinfoil covered jars. Heat syringe lift foil and inject.

I reheated the lighter after every injection and covered jars again and placed them individually inside zip lock bags. I sprayed the inside of the bags, including the covered jars again with bleach solution and sealed.

I sterilized a big tupperware container and put zip lock bags inside and sprayed again for good measure and closed lid. Put the tupperware inside a black bag and sprayed AGAIN.
All goes well for 5 days then I start seeing pink. I don't think there is anything else I can do other then investing in a sterile room...

Any ideas?


Edited by redrum (11/03/15 11:58 AM)


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OfflineDrCrumbs
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Re: Constant Contamination [Re: redrum]
    #22470746 - 11/03/15 12:04 PM (8 years, 2 months ago)

Which pf tek?

It sounds like you're going crazy with the chemicals. Spraying inside the PC is actually making it dirtier, since it was sterile before and is now just sanitized.

I dunno what you mean by spraying inside the bags.

What are you using to inoculate with?
This seems more likely where your issue is.

Oh I see your keeping them in bags. How do you keep them upright?
Also fresh air is your best Friend.


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Offlinemushpunx
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Re: Constant Contamination [Re: redrum]
    #22470764 - 11/03/15 12:08 PM (8 years, 2 months ago)

Sounds like a bad syringe man. If you are sterilizing your jars in pressure cooker you say? 15PSI for at least an hour and flame sterilizing the syringe between injections and still losing every jar its likely your syringe.




You're going way overkill though. Lose the bleach. Sterilize the jars and take the foil off.
No tubs or ziplock bags. That bleach under the foil might actually drip down inside your jars and cause contamination.

Make a still air box and inoculate them inside there. That's the only thing I would sugguest, that you read up on how to maje and properly use a still air box.

Honestly though if you flame sterilize your syringe then you can inoculate PF jars in open air without any problems.

Just make jars, sterilize, flame needle and inoculate in SAB and put them on a shelf.


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Edited by mushpunx (11/03/15 12:11 PM)


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Offlinemushpunx
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Re: Constant Contamination [Re: mushpunx]
    #22470791 - 11/03/15 12:13 PM (8 years, 2 months ago)

Are you buying the syringes or making them yourself? Are they coming from a sponsor?
Have you lost jars with more than one syringe?

And lastly are you using a dry vermiculite layer on your jars?


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OfflineArgyreia
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Re: Constant Contamination [Re: mushpunx]
    #22470807 - 11/03/15 12:15 PM (8 years, 2 months ago)

:whathesaid:

Yeah, you overkill. When doing the pf tek, you don't need alcohol. You can work in open air. Just take all your jars out of the PC and remove the foil when you are about to inject after having flamed the needle. After that, all the airborne contaminants will be trapped in the dry verm layer. No mask, no gloves, nothing needed.
If the syringes are an issue, you can do them yourself. It's foolproof and oable in open air too.


Edited by Argyreia (11/03/15 12:16 PM)


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OfflineDrCrumbs
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Re: Constant Contamination [Re: mushpunx]
    #22470810 - 11/03/15 12:16 PM (8 years, 2 months ago)

Yeah looking up a SAB will help you a lot. Even if you dont use it, but I would recommend it at this point. It may solve some unknown issue, like breathing on stuff, etc.

I just use a cleaned room, let to still when working from syringes.

Flame, then the needle touches nothing else before jar. Some squirt away first to cool.


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Invisibletahoe
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Re: Constant Contamination [Re: DrCrumbs]
    #22470913 - 11/03/15 12:37 PM (8 years, 2 months ago)

Sounds like a bad syringe. But who knows. I would not trust that one is following directions to a t if they are trying to sanitize their pc.


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Invisibleredrum
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Re: Constant Contamination [Re: tahoe]
    #22517394 - 11/13/15 09:49 AM (8 years, 2 months ago)

Thanks for the replies and sorry for such a late response. I was away on vacation :cool:

The first time I tried, I didn't use bleach or Lysol. I just sterilized the jars, flamed needle, and inoculated in open air. It was the second time I went overboard because I thought I was not careful the first round, but that obviously was not the case.

Both times, I used a dry layer of vermiculite on top. I used 2 separate syringes, both from the same sponsor on this site. Is it possible that BOTH separate syringes were contaminated?

According to the youtube video, "Lets grow Mushrooms", he sprayed down the PC as well before and after opening and right before closing the storage container for extra caution.


Edited by redrum (11/13/15 09:53 AM)


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Offlinemushpunx
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Re: Constant Contamination [Re: redrum]
    #22517455 - 11/13/15 10:11 AM (8 years, 2 months ago)

Yes its possible both syringes were bad but our sponsors are usually good enough for BRF cakes.

You dont wipe the needle or anything after flaming it right? Just get it glowing and shoot a lil liquid thru to cool it, then inoculate?


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Invisibleredrum
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Re: Constant Contamination [Re: mushpunx]
    #22517468 - 11/13/15 10:14 AM (8 years, 2 months ago)

Yes, that is correct Sir.


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Offlinemushpunx
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Re: Constant Contamination [Re: redrum]
    #22517472 - 11/13/15 10:15 AM (8 years, 2 months ago)

Run us thru how you are sterilizing the jars again please


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Invisibleredrum
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Re: Constant Contamination [Re: mushpunx]
    #22517499 - 11/13/15 10:24 AM (8 years, 2 months ago)

Quote:

mushpunx said:
Run us thru how you are sterilizing the jars again please




I start by cleaning out each jar and my PC with an antibacterial soap and warm water, rinsing well. I prepare my substrate. I spoon the substrate into the jars, leaving about an inch of space from the top. I then top with dry verm. I take a piece of foil, folded in half and cover the top of the jars, gripping around the edges to seal it as well as possible. I take another single layer of foil and place it on top of the first layer. I fill my PC with about 2 inches water and place the metal grate inside. Put the lid on and cook for 1hr at 15psi.

The second time, I did the exact same steps, except I used the lids of the mason jars with two holes, covered with a single layer of foil.

Both times, jars started going to shit after 5 days. ALL the jars.


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Offlinemushpunx
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Re: Constant Contamination [Re: redrum]
    #22517538 - 11/13/15 10:40 AM (8 years, 2 months ago)

Wow yea. If you are flaming your needke and the jars are PC' Im pretty certain its the syringes.

Since you have a PC you should try agar, maybe Pasty Plates first. You can isolate away from any contams


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Invisibleredrum
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Re: Constant Contamination [Re: mushpunx]
    #22517623 - 11/13/15 11:09 AM (8 years, 2 months ago)

Quote:

mushpunx said:
Wow yea. If you are flaming your needke and the jars are PC' Im pretty certain its the syringes.

Since you have a PC you should try agar, maybe Pasty Plates first. You can isolate away from any contams



Hahah your speaking another language. Care to point me in the right direction? Is this method easy enough for beginners?


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InvisibleShroomspective
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Re: Constant Contamination [Re: redrum]
    #22517638 - 11/13/15 11:13 AM (8 years, 2 months ago)



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Invisibleredrum
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Re: Constant Contamination [Re: Shroomspective]
    #22518305 - 11/13/15 02:17 PM (8 years, 2 months ago)

Quote:

Shroomspective said:
Pastywhyte's Easy Agar Tek :thumbup:




Thank you for that. Looks intimidating. I might give the PF tek another go with a fresh syringe.


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OfflineDrCrumbs
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Re: Constant Contamination [Re: redrum]
    #22518362 - 11/13/15 02:32 PM (8 years, 2 months ago)

Its not though it actually makes things way better and with much less wasted work.


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Offlinemushroomnoob1981
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Re: Constant Contamination [Re: DrCrumbs]
    #22521375 - 11/14/15 08:26 AM (8 years, 2 months ago)

A mistake I've made = letting bubbles of air into the spore syringe.  You or your vendor may have used sterile water to make the syringe, but air isn't sterile unless it's fresh out of a PC/autoclave or just passed through a HEPA filter. 

Also, if you're looking for a chemical that will kill off anything... 1 M sodium hydroxide + 10% bleach.  Warning: this stuff is incredibly caustic and will burn your skin/clothes... god help you if you put it in a spray bottle.  It will reduce all mold/bacterial spores to ash, though.


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Offlinethebug76
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Re: Constant Contamination [Re: mushroomnoob1981]
    #22521409 - 11/14/15 08:42 AM (8 years, 2 months ago)

TIP: Keep the needle pointed up while flaming, don't pull any air into the syringe, make sure it gets red hot, don't wipe it with anything, don't put the foil back on after inoculation.

PF tek is all I've done so far with colonization, but I am yet to see a mold in a jar and I only steam because I don't have a PC. A little bacteria a couple times tho.


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Offlinemushpunx
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Re: Constant Contamination [Re: thebug76]
    #22521538 - 11/14/15 09:22 AM (8 years, 2 months ago)

Then what are the trays in your sig, that's PF tek but in like PP5s? They are gorgeous


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Offlinethebug76
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Re: Constant Contamination [Re: mushpunx]
    #22522012 - 11/14/15 11:38 AM (8 years, 2 months ago)

I did PF tek, then crumbled and layered with hpoo/CVG in a mini tray setup. I haven't fruited straight cakes tho.

I've just been pretty lucky with the jars by following the tips, I gave earlier, and they've all been MS to PF jars.


Edited by thebug76 (11/14/15 11:41 AM)


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OfflinecronicrFacebook
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Re: Constant Contamination [Re: thebug76]
    #22522261 - 11/14/15 12:40 PM (8 years, 2 months ago)

pink mold is dirty inoculant or lack of sterile procedure


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OfflinePsilosoulful

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Re: Constant Contamination [Re: redrum]
    #22522473 - 11/14/15 01:33 PM (8 years, 2 months ago)

Quote:

redrum said:
Hey guys,

I tried twice to cultivate following pf tek to a "T". Pink mold is a constant issue.

Washing the jars with antibacterial soap, spray with bleach, and rinse again. Prepare the substrate and in the cooker they go. 15psi for 1hr.



Try PC'ing for 90 minutes.


Quote:

redrum said:
Pot cools down for about 6 hours. Prepare room by spraying EVERYTHING down with bleach and lysol, including pressure cooker before opening and letting it sit for 10 min. I even wore rubber gloves and sprayed them. I opened the cooker and sprayed bleach solution inside on tinfoil covered jars. Heat syringe lift foil and inject.

I reheated the lighter after every injection and covered jars again and placed them individually inside zip lock bags. I sprayed the inside of the bags, including the covered jars again with bleach solution and sealed.

I sterilized a big tupperware container and put zip lock bags inside and sprayed again for good measure and closed lid. Put the tupperware inside a black bag and sprayed AGAIN.
All goes well for 5 days then I start seeing pink. I don't think there is anything else I can do other then investing in a sterile room...

Any ideas?



You need to build a SAB to do your inoculation work. It's super simple and very effective. I rarely get contams when using proper sterile techniques. Get a 66qt tote or larger and melt two large holes big enough for your hands to fit through on one of the long sides. Additionally, you can attach some tyvek sleeves to the outside.


And check out the first link in my sig. :thumbup:


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Offlinethebug76
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Re: Constant Contamination [Re: mushpunx]
    #22525188 - 11/14/15 10:18 PM (8 years, 2 months ago)

Quote:

mushpunx said:
Then what are the trays in your sig, that's PF tek but in like PP5s? They are gorgeous




Oh yea, thanks by the way. I was really proud of those, especially since it was my first successful grow. Sorry I forgot to say thanks earlier, I was in a hurry getting ready for work. I've got another grow in progress. Still in the jar stage now, but I'm planning 12 - 1qt pp5's instead of 3 like that first grow and they're all gonna be the bigger strains (6 GT and 6 AA+).


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Edited by thebug76 (11/14/15 10:19 PM)


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Invisibleredrum
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Re: Constant Contamination [Re: thebug76]
    #22526782 - 11/15/15 09:19 AM (8 years, 2 months ago)

That might might have been part of the problem. There was no air in the syringe for me to shake it to break up the clumps so I pulled back the plunger while the needle was in fire to ensure the air was sterile.

I just remembered I tossed all my jars after being annoyed at the contamination. All I have now is pint size jars. Are they too big for pf tek?


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Invisibleredrum
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Re: Constant Contamination [Re: thebug76]
    #22526805 - 11/15/15 09:24 AM (8 years, 2 months ago)

Quote:

thebug76 said:
TIP: Keep the needle pointed up while flaming, don't pull any air into the syringe, make sure it gets red hot, don't wipe it with anything, don't put the foil back on after inoculation.

PF tek is all I've done so far with colonization, but I am yet to see a mold in a jar and I only steam because I don't have a PC. A little bacteria a couple times tho.



Don't put foil back in after inoculation? I'm going to have two tiny open holes in the bottom layers of foil. Won't that put the jars more at risk?


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Offlinethebug76
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Re: Constant Contamination [Re: redrum]
    #22526880 - 11/15/15 09:55 AM (8 years, 2 months ago)

Quote:

redrum said:
Quote:

thebug76 said:
TIP: Keep the needle pointed up while flaming, don't pull any air into the syringe, make sure it gets red hot, don't wipe it with anything, don't put the foil back on after inoculation.

PF tek is all I've done so far with colonization, but I am yet to see a mold in a jar and I only steam because I don't have a PC. A little bacteria a couple times tho.



Don't put foil back in after inoculation? I'm going to have two tiny open holes in the bottom layers of foil. Won't that put the jars more at risk?




When your using PF tek, the dry verm acts as a barrier and leaving tge foil off allows the tops of the jars to dry out. Leaving it on, traps moisture which germinates contams and lets the moisture, as well as the contam, to work its way thru the verm barrier to your sub.


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Invisibleredrum
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Re: Constant Contamination [Re: thebug76]
    #22527171 - 11/15/15 11:36 AM (8 years, 2 months ago)

When your using PF tek, the dry verm acts as a barrier and leaving tge foil off allows the tops of the jars to dry out. Leaving it on, traps moisture which germinates contams and lets the moisture, as well as the contam, to work its way thru the verm barrier to your sub.



Interesting. Every thing I read and watched insists on putting the foil back on. I do see your point and it makes sense. I will leave the top layer off this time and just leave the layer I inoculate through on.
Thanks

Once inoculated, can I put them in a plastic container then in a black garbage bag to keep them dark, or just leave them out in open air?
Can I use pint size jars and cut them in half once birthed?


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Offlinethebug76
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Re: Constant Contamination [Re: redrum]
    #22527275 - 11/15/15 12:05 PM (8 years, 2 months ago)

I just put them in a clothes drawer that has a little extra room, but most of the guys on here say just stick them on a shelf. Light doesn't hurt anything and you want room temperature.


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Offlinethebug76
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Re: Constant Contamination [Re: thebug76]
    #22527350 - 11/15/15 12:26 PM (8 years, 2 months ago)

I wouldn't use anything bigger than 1/2 pint (8 oz) for PF tek, otherwise they tend to stall out. And yes open air.


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