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Offlinewedder
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Need help identifying contams (pics)
    #12567607 - 05/15/10 02:33 PM (14 years, 8 months ago)

I incubated these jars 2 weeks ago. I have noticed a small discoloration on the mycelium in a few jars. I need help identifying if these jars are contaminated.




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OfflineStainBlue
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Re: Need help identifying contams (pics) [Re: wedder]
    #12567613 - 05/15/10 02:35 PM (14 years, 8 months ago)

those pics are a little blurry it's hard to tell.

Maybe try posting this over in the contamination forum.


--------------------
Peace,
StainBlue

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Offlinebw86
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Re: Need help identifying contams (pics) [Re: wedder]
    #12567614 - 05/15/10 02:35 PM (14 years, 8 months ago)

NEED HELP FINDING CONTAM FORUM
edit- why would you want to identify your mold? are you trying to isolate a specific type?

Edited by bw86 (05/15/10 02:37 PM)

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Invisiblegeorge castanzaM
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Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12567619 - 05/15/10 02:37 PM (14 years, 8 months ago)

This thread was moved from Mushroom Cultivation.

Reason:
More/better responses in the proper forum.

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Offlinewedder
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Re: Need help identifying contams (pics) (moved) [Re: george castanza]
    #12567684 - 05/15/10 02:55 PM (14 years, 8 months ago)

Sorry I posted this in the wrong forum. Thanks for moving it.

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Offlinepairshroom
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Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12567990 - 05/15/10 03:52 PM (14 years, 8 months ago)

These pics are a bit dark. You might try reshooting the photos with more ambient light.

Unfortunately, it looks like a green contaminant on your colonizing jars. Usually, this will be Trichoderma spp. (about 35 or so different species), the forest green mold. It is an unusually aggressive, and often parasitic mold, which invariably spells doom for growing mycelium, such as Psilocybe. The spores of this fungus are green and incredibly "sticky" and robust. Once released into your living environment, it can be nearly impossible to eradicate it, jeopardizing future grow attempts.

What this means is that you should toss the jars (don't open them), if possible. If you need to keep them, you should sterilize them the jars in a PC first and then dispose of them. Overall, sometimes it's easier to just toss the whole jar and replace it with a new one(s).

The reason for the Trich usually relates to suboptimal sterile technique. You might wish to review your tek, and try it again paying attention to this important aspect.

Edited by pairshroom (05/16/10 07:35 AM)

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Offlinewedder
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Re: Need help identifying contams (pics) (moved) [Re: pairshroom]
    #12570559 - 05/15/10 11:58 PM (14 years, 8 months ago)

I have all my jars next to each other, all of which have their lids on but there is four holes in each lid. Each jar has 1 inch of dry verm on the top. Could this mold transfer from one jar to another?

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Offlinepairshroom
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Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12571729 - 05/16/10 06:41 AM (14 years, 8 months ago)

Assuming the holes are relatively small, cross-contamination would be unlikely. Also, the dry verm barrier at the top is pretty effective at keeping mold spores out. This would suggest that contamination occurred because (a) the jars weren't properly sterilized, (b) the syringe was contaminated, or (c) contamination was introduced during inoculation due to improper technique.


--------------------
"Beware the Jabberwock, my son!
  The jaws that bite, the claws that catch!
Beware the Jubjub bird, and shun
  The frumious Bandersnatch!"

— L. Carroll

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Offlinewedder
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Re: Need help identifying contams (pics) (moved) [Re: pairshroom]
    #12573425 - 05/16/10 02:16 PM (14 years, 8 months ago)

Here are some better pictures.




I had one person tell me it looks like a yeast infection and another person tell me it is a contaminant. I hope it isn't anything serious because it's is showing up in half of my jars.

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Offlinepairshroom
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Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12574025 - 05/16/10 04:07 PM (14 years, 8 months ago)

I don't think this is yeast, which is usually colorless or white, and certainly not green. Yeast does not generate spores, and it is the spores (which occur in molds) that have the color (in this case green).

The new pics are highly suspicious for Trichoderma (aka Trich). I would start over at this point. Read up a bit more on BRF PF TEK on this site, and make sure you take every precaution with being clean/sterile. If you open these jars, you will jeopardize any future grow attempts, despite being more clean.

Better luck next time.


--------------------
"Beware the Jabberwock, my son!
  The jaws that bite, the claws that catch!
Beware the Jubjub bird, and shun
  The frumious Bandersnatch!"

— L. Carroll

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Offlinewedder
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Re: Need help identifying contams (pics) (moved) [Re: pairshroom]
    #12574045 - 05/16/10 04:09 PM (14 years, 8 months ago)

Well that really sucks. Too bad, it is contaminated over half of my jars. I thought I was being very sterile when I first did this but I guess it wasn't enough. Thanks for your help.

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Offlinepairshroom
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Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12574121 - 05/16/10 04:24 PM (14 years, 8 months ago)

You'll get it right. Just keep trying. Review your Tek and think where contamination might have occurred.

(1) spores - did you get them from a Shroomery sponsor?
(2) jars - did you wipe and dry the edges of the jars before putting the lids on?
(3) jars - did you cover with Al foil when sterilizing?
(4) jars - did you add a 1/2" dry verm layer?
(5) jars - did you PC for 90 mins at 15 psi?
(6) jars - how big are the holes in the lid (1/6" is about right)?
(7) inoculation - did you shower, brush teeth, wear gloves, mask, hat, gown (believe it or not, I do all of this)?
(8) inoculation - did you use a glove box or similar? did you wipe it with alcohol first? did you use a room without moving air?
(9) inoculation - did you flame sterilize your needle between jars?


If you haven't already, review RogerRabbit's Tek at www.mushroomvideos.com. It has just about all you need.


--------------------
"Beware the Jabberwock, my son!
  The jaws that bite, the claws that catch!
Beware the Jubjub bird, and shun
  The frumious Bandersnatch!"

— L. Carroll

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Offlinewedder
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Registered: 04/24/10
Posts: 46
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Re: Need help identifying contams (pics) (moved) [Re: pairshroom]
    #12574667 - 05/16/10 05:58 PM (14 years, 8 months ago)

I did do 1-4.
Number 5, instead of using a pressure cooker I used a regular pot for 120 minutes. I hope that isn't where the contam came from. I did read on this website in multiple places that not using a pressure cooker shouldn't make a difference.
Regarding number 7 and 8, I misted bleach throughout my bathroom twice. During incubation I wore rubber Glove and I had a T-Shirt tied around my face. But I did not shower right before and I did not wear a hat so my hair was exposed.
I did flame the needle each time, but I used a Bic lighter which leaves black residue. I also read on here somewhere that the black residue doesn't make any difference.
Oh and I got my syringe from ralph spores.

Edited by wedder (05/16/10 06:08 PM)

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Offlinewedder
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Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12574688 - 05/16/10 06:01 PM (14 years, 8 months ago)

So I incubated a total of 8 jars. Of the 8 I suspect 7 have contams. Hear is a picture of each jar where I suspect contamination. (I am sorry the pictures may not be the best of quality.)

1

2

3

4

5

6

7

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Offlinepairshroom
Jabberwocky
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Registered: 01/01/10
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Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12574945 - 05/16/10 06:46 PM (14 years, 8 months ago)

A pressure cooker is ideal, but prolonged heating in boiling water is a valid tek, as well. The reason a pressure cooker is better is that it can achieve higher temperatures. Boiling water will reach a maximum of 212F (100C) - steam then develops, so the max liquid temp is that of boiling water.  Under pressure, however, boiling/vaporization occurs at a higher temp of 252F (122C). While molds and most bacteria are killed at 212F, some bacterial spores (e.g.,. Clostridium botulinum) are resistant and require higher temps of 240F. You can compensate, however, using prolonged boiling at 90-120 mins (PC requires only about 45 mins once 15 psi is reached).

Hair is a prominent source of bacteria and molds, so a disposable surgical hat is important, as is clean, washed hair. You can get them cheaply at medical supply stores or online. You can also use a clean painter's hat from a home center or paint store.

I would not inoculate in the bathroom. Of all of the rooms in the house, bathrooms have the highest mold levels. Choose a clean, dry (not damp) area in the house, and convert a clear Sterilite or Rubbermaid tote into a clean glove box. Use a surgical gown or at least Tyvek sleeves (found at Lowes or paint stores, too), to prevent contamination from arm hair and dead skin. Use gloves, and wipe down with alcohol frequently.

Basically, you can't be too clean when performing this critical step. Once your cakes fully colonize and you birth them, they are relatively resistant to contamination.

I don't personally deal with Ralphster's, but many do, and I have never heard about any issues, so you're probably OK in this regard.

The carbon residue from a Bic lighter is inert, so that's OK. However, using a lighter takes one hand, and you might lose the sterility of your freshly alcohol-wiped gloves. I wouldn't want to light a Bic lighter with alcohol on my hands anyway! Just invest in an alcohol lamp for $5 (http://www.amazon.com/ALCOHOL-LAMP-BURNER-Flint-glass/dp/B0018BJW6S/ref=sr_1_1?ie=UTF8&s=home-garden&qid=1274056840&sr=1-1 or check eBay, etc.). It's faster, easier, and cleaner (just remember the flame can be nearly invisible, so don't get faked out).

Also, you appear to be using regular 1/2 pint mason jars, which are OK. I prefer the wide mouth 1/2 pint jars (Jarden/Kerr), which are more squat and I find they tend to colonize a bit faster. Jarden/Ball doesn't make them anymore, so they can be tough to find, but I found a bunch at Ace Hardware, and got others online (http://www.amazon.com/gp/product/B0000BYD0F/ref=s9_simh_gw_p79_i1?pf_rd_m=ATVPDKIKX0DER&pf_rd_s=center-2&pf_rd_r=11EJ94CQ24VBETZRF90E&pf_rd_t=101&pf_rd_p=470938631&pf_rd_i=507846 or check eBay, etc.).

Review RR's PF Tek on his site or better yet, buy his DVD. It's an excellent reference not only for PF TEK, but for most of the advanced techniques you will likely ultimately graduate upto.


--------------------
"Beware the Jabberwock, my son!
  The jaws that bite, the claws that catch!
Beware the Jubjub bird, and shun
  The frumious Bandersnatch!"

— L. Carroll

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Offlinewedder
First-Time Shroomer
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Registered: 04/24/10
Posts: 46
Last seen: 14 years, 6 months
Re: Need help identifying contams (pics) (moved) [Re: pairshroom]
    #12575257 - 05/16/10 07:53 PM (14 years, 8 months ago)

Well, I have one jar which isn't contaminated. I don't mind redoing everything, but I don't have any spores. Is there a way I can get spores from my clean jar and spread them throughout out many jars?

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Offlinepairshroom
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Registered: 01/01/10
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Last seen: 13 years, 7 months
Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12575612 - 05/16/10 09:00 PM (14 years, 8 months ago)

No. You need mushrooms to get spores. You can recover them from mycelia, and even if you could, I wouldn't risk using the 1-out-of-8 jars you assume is uncontaminated. It probably is, and it hasn't declared itself yet. Remember that Trich mycelium is white, too - the green only occurs once sporulation occurs.

I would get a new spore syringe.


--------------------
"Beware the Jabberwock, my son!
  The jaws that bite, the claws that catch!
Beware the Jubjub bird, and shun
  The frumious Bandersnatch!"

— L. Carroll

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Offlinewedder
First-Time Shroomer
Male

Registered: 04/24/10
Posts: 46
Last seen: 14 years, 6 months
Re: Need help identifying contams (pics) (moved) [Re: pairshroom]
    #12575970 - 05/16/10 10:07 PM (14 years, 8 months ago)

Oh well what a shame. I'm not looking forward to dropping another $20 to get another syringe.

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Offlinepairshroom
Jabberwocky
Male


Registered: 01/01/10
Posts: 57
Last seen: 13 years, 7 months
Re: Need help identifying contams (pics) (moved) [Re: wedder]
    #12577921 - 05/17/10 09:03 AM (14 years, 8 months ago)

Call or e-mail the company. They might make a concession (e.g., discounted replacement) if you explain that your jars contaminated.


--------------------
"Beware the Jabberwock, my son!
  The jaws that bite, the claws that catch!
Beware the Jubjub bird, and shun
  The frumious Bandersnatch!"

— L. Carroll

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Offlinewedder
First-Time Shroomer
Male

Registered: 04/24/10
Posts: 46
Last seen: 14 years, 6 months
Re: Need help identifying contams (pics) (moved) [Re: pairshroom]
    #12580368 - 05/17/10 04:57 PM (14 years, 8 months ago)

Call me crazy, but I made an attempt to get rid of the contaminations. I used a sterile spoon in a sterile room and dug out the contamination as well as some of the mycelium which was close to the contamination. I don't know if this will work but it will be a miracle if it does.

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