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Ca_Gone
Stranger


Registered: 03/04/12
Posts: 12
Loc: Australia
Last seen: 3 years, 7 months
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Help, reoccurring purple contam
#26619342 - 04/22/20 05:33 AM (3 years, 9 months ago) |
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Hey guys I apologize for the long post I just need to know if I'm doing something wrong. I've had my last 2 pf tek attempts fail with the same purple contamination. 1st attempt 10brf jars steamed for 2 hours, allowed to cool overnight in pot, work bench, SAB and equpiment wiped down with 70% iso, jars loaded in SAB and sprayed again with 70%iso. Waited 5min for vapour to disperse. Needle heated outside of SAB until red Hot, brought inside SAB wiped with Iso (I know my bad) then removed foil on 1 jar inoculated with GT spore syringe then a 2nd jar before reheating needle. All jars 100% colonized in 10 days but 70% of the jars started to show purple hues at the inoculation point.
I decided to bin them all. Jars opened outside cakes binned then soaked jars in 10% bleach for 12hrs before bringing inside washed with soapywater and baked jars for 1.5hrs at 180°C (356°F) in oven.
2nd attempt repeated exactly the same however the spore syringe was from a different vendor and I didnt wipe the needle with iso after flame sterilizing. 2nd batch came out worse, all 12 jars and at all 4 innoc points showed purple contam at 7th day. 2 of those jars had never been used, steamed by themselves for 2 hours and inoculated on a separate day with a new needle (same syringe) and still had the same contam, no other type of contam was observed on any jar on either attempt and all the purple was at the inoculation points. I've bought a PC now but it will be awhile til its delivered, I also plan on getting into agar.
Any tips on what I might of screwed up and is any of the white growth, mushroom mycelium or it all just part of the purple contam? I took a break for 7yrs so cant remember exactly what mycelium should look like.
 First attempt, worst jar day 11 after inoculation.
2nd attempt pic typical of all jars, day 7 after inoculation.
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rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
Posts: 6,843
Loc: In the Gills
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Re: Help, reoccurring purple contam [Re: Ca_Gone] 1
#26619423 - 04/22/20 07:06 AM (3 years, 9 months ago) |
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Seems how all of your jars failed both times with two different syringes and not one made it. I'm gonna say maybe it's your 2hr steam that's just not getting them sterile maybe. I know the tek says it works but I'd still rather use a pressure cooker to be sure.
Getting into agar is definitely a better way to germinate spores and you'll need a pressure cooker for that anyhow.
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LogicaL Chaos
Ascension Energy & Alien UFOs




Registered: 05/12/07
Posts: 69,599
Loc: The Inexpressible...
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Re: Help, reoccurring purple contam [Re: Ca_Gone]
#26619453 - 04/22/20 07:27 AM (3 years, 9 months ago) |
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That is really weird. Almost looks like spores. Similar color. Never seen anything like it....
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Tallman
Mycophile



Registered: 03/25/20
Posts: 27
Loc: Göbekli Tepe
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Definitely use your PC! Be patient!! Steam works but 2hrs is definitely the problem IMO
-------------------- MushLove
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Ca_Gone
Stranger


Registered: 03/04/12
Posts: 12
Loc: Australia
Last seen: 3 years, 7 months
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Re: Help, reoccurring purple contam [Re: Tallman]
#26620737 - 04/22/20 06:03 PM (3 years, 9 months ago) |
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Thanks for the replies. Steam worked for me 7 yrs ago. Only ever encountered trich back then but this contam is confusing me trying to figure out the source.. syringe, substrate etc.? I'll hold off on the 3rd attempt until I recieve my PC so I can rule out substrate sterilization, thanks again.
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poisoned
untitled



Registered: 04/17/13
Posts: 1,738
Loc: Yurop
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Re: Help, reoccurring purple contam [Re: Ca_Gone] 1
#26621436 - 04/23/20 03:31 AM (3 years, 9 months ago) |
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PF tek is working 99% of the time with steam, that's the whole purpose of the tek.
Do you have a dry verm layer? If you do, maybe try longer steam times. You're flaming the needle before inoculating tho, right?
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Ca_Gone
Stranger


Registered: 03/04/12
Posts: 12
Loc: Australia
Last seen: 3 years, 7 months
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Re: Help, reoccurring purple contam [Re: poisoned]
#26623546 - 04/23/20 10:04 PM (3 years, 9 months ago) |
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Yeah if you look at the pics where that first ridge is on the jar, I fill brf up to there, wipe the remaining part of jar with a dry paper towel and fill with dry verm. I flame sterilize after every 2nd jar but might try after each jar. Might try soapy water spray in SAB with a towel on the bottom wet with 10% bleach. Instead of 70% ISO I spray only once at the start because I'm Worried about to much iso vapour close to an ignition source.
For argument sake let's say the 2 hours for the 10 half pint jar wasnt enough. Surely the 2 jars by themselves for 2 hours was enough but they had the same outcome. Also If the jars werent sterilized properly wouldnt the contam be in random spots, 100% of contam has been at inoculation points.
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