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 vnc
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Registered: 10/22/10
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Last seen: 13 years, 5 months
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 9 jars, all contaminated differently 
#13370530 - 10/22/10 09:48 AM (13 years, 5 months ago) |
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Hello,
it's my first post here about my first try to grow shrooms from the spores.
After sterilizing 9 jars I put a lot of spores in there, used 3 syringes and 9 needles (1 needle per jar, 1 syrynge for 3 jars). Now, 10 days later, I see in every single jar something that does not belong there.
There are 4 colors of contams: black, red, orange/green and blue/green. Every jar seems to have catched its own contam.
I suppose I can stop the experiment and throw everything away, but how can I know, at what stage something went wrong? As far as I can tell, I did everything exactly as explained in PF-TEK videos/instructions.
The only reason I'm adding a new thread is that maybe the existance of that many kinds of mould is a hint on what happened.
Thanks! Vnc
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baraka
 
Registered: 07/15/00
Posts: 10,768
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Re: 9 jars, all contaminated differently [Re: vnc]
#13370703 - 10/22/10 10:39 AM (13 years, 5 months ago) |
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Poor sterilization of the substrate is where you went wrong most likely. Getting a pressure cooker is ideal. If you are boiling jars not under pressure, they need to have a steady boil covered for at least and hour. Another possible problem is you had too much water in the jars.
Also you do not need to overdo putting spores in jars. A little goes a long way. When doing half pint jars i used to do roughly a half to two thirds of a syringe for 12 jars. Once you see liquid coming out of the syringe into the inoculation point you can stop.
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This is the only time I really feel alive.
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vnc
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Re: 9 jars, all contaminated differently [Re: baraka]
#13371095 - 10/22/10 11:57 AM (13 years, 5 months ago) |
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Quote:
Poor sterilization of the substrate is where you went wrong most likely. Getting a pressure cooker is ideal. If you are boiling jars not under pressure, they need to have a steady boil covered for at least and hour.
boiled them for 90 minutes
Quote:
Another possible problem is you had too much water in the jars.
don't think so
Quote:
When doing half pint jars i used to do roughly a half to two thirds of a syringe for 12 jars. Once you see liquid coming out of the syringe into the inoculation point you can stop.
That's what I did definitely wrong. I put 3ml of distilled water with spores into every jar. Can this be the reason?
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dizzyease
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Re: 9 jars, all contaminated differently [Re: vnc]
#13371135 - 10/22/10 12:06 PM (13 years, 5 months ago) |
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id say yes. i get bac contams if i use too much water and i boil for ninety minutes aswell
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baraka
Registered: 07/15/00
Posts: 10,768
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Re: 9 jars, all contaminated differently [Re: vnc]
#13371328 - 10/22/10 12:58 PM (13 years, 5 months ago) |
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Quote:
vnc said:
Quote:
Poor sterilization of the substrate is where you went wrong most likely. Getting a pressure cooker is ideal. If you are boiling jars not under pressure, they need to have a steady boil covered for at least and hour.
boiled them for 90 minutes
If you had a wide variety of different colors then the sterilization had to of been not done well enough. If it came from the syringe you would likely have just one type of contam, most likely a bacteria.
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This is the only time I really feel alive.
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saint_peter
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Re: 9 jars, all contaminated differently [Re: baraka]
#13375016 - 10/23/10 07:09 AM (13 years, 5 months ago) |
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Hmm, I wonder if changing needles inbetween jars does more bad than good...
How do you keep the joint between syringe and needle sterile during the needle change?
I think proper flame-sterilisation until the tip of the needle glows red, every time before you put the needle into a jar, is a safer way to keep out contams. Not even the hardiest bacterial endospores can survive 400-500°C, and you can keep the time between cooling of the needle and insertion into the jar reasonably short.
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baraka
Registered: 07/15/00
Posts: 10,768
Loc: hyperspace
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Re: 9 jars, all contaminated differently [Re: saint_peter]
#13375306 - 10/23/10 09:18 AM (13 years, 5 months ago) |
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When i used to do jars i would just wipe the needle down with iso alcohol and shoot up 12 jars with out stopping or cleaning the needle again. I would almost never have contamination problems and if i did i would get a bacteria i would attribute to improper water content. Some times get extra water in the jar when pressure cooking.
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This is the only time I really feel alive.
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vnc
Stranger
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Re: 9 jars, all contaminated differently [Re: saint_peter]
#13379058 - 10/24/10 03:41 AM (13 years, 5 months ago) |
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Quote:
saint_peter said:
I think proper flame-sterilisation until the tip of the needle glows red, every time before you put the needle into a jar, is a safer way to keep out contams.
Ok, I'll try that next time
BTW, here is the pic:
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baraka
Registered: 07/15/00
Posts: 10,768
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Re: 9 jars, all contaminated differently [Re: vnc]
#13379555 - 10/24/10 09:33 AM (13 years, 5 months ago) |
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Yikes.
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This is the only time I really feel alive.
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 Mr. Anderson
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Registered: 09/05/10
Posts: 2,677
Loc: Torn between the roots of...
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Re: 9 jars, all contaminated differently [Re: baraka]
#13379593 - 10/24/10 09:47 AM (13 years, 5 months ago) |
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Be careful with the black mold, it can be toxic. U wanna pc or boil ur jars for a couple hrs just to make sure its dead before opening.
For the green trich contam u can probably just open as is OUTSIDE and burry them,most of times they will still flush. If u do this id wear a respirator or even a sheet tied around ur mouth and nose and where some clothes u can dispose of, old stuff. Then hop in the shower after
Changing the needles that much sounds like ur asking for trouble, just follow the tek as is. I just swab the needle with 70% alcohol then take the needle to the outside of my glovebox to flame steralize with an already lit alcohol lamp, this just needs to be done in between jars.
Did u use a glove box and those jars look a little big, u should use half pint wide mouth jars. Regular mouth or taller jars can be more prone to ur verm barrier being compromised by sinking and falling down wen ur substrate shrinks.
Also just wondering did u incubate? If the jars are at higher temps it can cause contams to grow fast if they're present.70-75 is good.
To me the problem sounds like the excessive switching of the needles.
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  Disclaimer: All posts are completely fictional and or for educational purposes only.
Edited by Mr. Anderson (10/24/10 09:52 AM)
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vnc
Stranger
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Re: 9 jars, all contaminated differently [Re: Mr. Anderson]
#13380780 - 10/24/10 03:33 PM (13 years, 5 months ago) |
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Thanks a lot for your answers!
Quote:
nutsmcbo said:
Be careful with the black mold, it can be toxic. U wanna pc or boil ur jars for a couple hrs just to make sure its dead before opening.
that's what I did with all of the jars at once after taking the picture.
Quote:
nutsmcbo said:
For the green trich contam u can probably just open as is OUTSIDE and burry them,most of times they will still flush.
had not expected anything to be able to flush yet, so I killed everything.
Quote:
nutsmcbo said:
Did u use a glove box and those jars look a little big, u should use half pint wide mouth jars.
they a littlemore than half a pint (300ml. 1/2pint=240ml)
Quote:
nutsmcbo said:
Also just wondering did u incubate? If the jars are at higher temps it can cause contams to grow fast if they're present.70-75 is good.
the temperature was not absolutely constant: 70-75 at night, up to 80 daytime, but never more than that.
Quote:
nutsmcbo said:
To me the problem sounds like the excessive switching of the needles.
Ok, I'll try with the flame next time, but I still don't get how new sterile needles can cause a problem.
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Mr. Anderson
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Registered: 09/05/10
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Loc: Torn between the roots of...
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Re: 9 jars, all contaminated differently [Re: vnc]
#13380985 - 10/24/10 04:15 PM (13 years, 5 months ago) |
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Quote:
vnc said: Ok, I'll try with the flame next time, but I still don't get how new sterile needles can cause a problem.
It's not the needles or new needles causing the problem. It's when ur taking them off to put the new ones on you are exposing the multispore solution at the end of the syringe to possible contams in the air. Thats why I only do this once and always use a glovebox. In between jars swab the needle with alcohol then flame steralize the tip of the needle til red hot outside the glovebox. If ur flame is inside the glovebox that is bad, heat from the flame will make air circulate defeating the whole purpose of the glovebox.
You can have a good success rate with innoculating ms when using a glovebox. The whole goal is to create a still air atmosphere to work in, it is not sterile by any means.
Just do it this way. After ur jars cool put them strait in ur glove box from the pc when you open it. Clean the surface where ur glove box will be sitting on before hand, if you use alcohol to clean the surface let the fumes evaporate first. It can cause some disturbance in the air if you don't, just don't leave a shit load of it.
Now just take the tinfoil off ur jars inside ur glovebox, all of them. Get ur syringe ready, swab with alcohol then stick it out the side of ur glovebox and flame sterilize it on ur alcohol lamp that should be lit and stays lit. Inoculate all 4 holes and then when you move to the next jar swab again with alcohol and flame steralize the tip out the side of the gb again.
I've been having 100 % success rate lately doing this.
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Disclaimer: All posts are completely fictional and or for educational purposes only.
Edited by Mr. Anderson (10/24/10 04:30 PM)
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