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Offlinefiegtree
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Registered: 10/04/15
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Why am I getting contaminants?
    #22414268 - 10/21/15 03:47 PM (8 years, 3 months ago)

To get right into it, I recently purchased 12 jars from a shroom kit company. I know this is frowned upon but I did it to be safe as its one of my first times. I was very careful with my inoculation. I warmed up the oven, pulled out the rack and so on. Used a cigarette lighter and alcohol towelette repetitively. I then placed them in my colonization chamber with the low air temp being 76 degrees, and the high air temp of 80 degrees. They were colonizing with 6 other jars that were a few weeks ahead. Near the top of a lot of the new 12 jars is this dark black region. I assume its contamination but if there is any other source for that kind of color I'll be more than happy to hear it. In some of the jars its much more dark and obvious than others but I feel like I can see feint black blobs on most if not all of the 12 jars. When I purchased 6 jars from the same company earlier, I only lost 1 to contamination. I'd post pics but it doesn't show too well. The black area is always near the top of the jar though. So I pulled about 5 of the worst looking jars out as I didn't want to contaminate any others. So besides the fact that the shroom kit company didn't sterilize properly, what are some sources of contamination? My second question is how possible is cross contamination in my described circumstances? Should I pull out every single suspicious jar, or can I leave them with the good ones in my enclosed chamber?


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OfflineYaMoonSun
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Re: Why am I getting contaminants? [Re: fiegtree]
    #22414304 - 10/21/15 03:57 PM (8 years, 3 months ago)

Quote:

Used a cigarette lighter and alcohol towelette repetitively.



Well, if you wiped the syringe with alcohol after flame sterilizing that could be your problem right there. The flame sterilizes, where as the alcohol only sanitizes, meaning that you're basically whipping the needle with a bunch of active mold spores after killing the other ones. :shrug:


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InvisiblePanda89
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Re: Why am I getting contaminants? [Re: fiegtree]
    #22414312 - 10/21/15 03:59 PM (8 years, 3 months ago)

Hey bro I am speaking from experience. I got jars from a outfit like you and they included instructions to do inoculation using the oven. After failing hard as fuck, I started reading more of the forums. The pf tek is super easy no pc required.the oven tek is outdated and does more harm then good. For future inoculations build a still air box you can't go wrong for a few bucks. Usually room Temps are good for colonization if your comfortable your shrooms are comfortable.


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OfflineSivarted
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Re: Why am I getting contaminants? [Re: Panda89]
    #22414381 - 10/21/15 04:18 PM (8 years, 3 months ago)

I never trusted the ideas behind the oven trick (although the friend that introduced ed me to this hobby swears by it), and my SAB is small enough that it's kind of a pain to work in, so when I'm inoculating pftek jars or grain jars with polyfill lids, I don't do either.  I just clean all the surfaces in a small bathroom and do it all there, with a pretty good success rate (I've only had 2 jars contaminate this way, out of somewhere around 30ish)

If *all* of your jars contaminated, either your syringe has contams, the batch of jars came contaminated, or you're wiping after flaming and introducing them that way.  As far as I know, there's no reason to ever use alcohol on a red hot flamed syringe.

I know the oven thing *can* work though, because that's how my friend does all his inoculations, so it would be weird for all of them to contam just because of the method, unless it creates weird air currents in your kitchen that shove everything back into the working area.


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InvisibleMad Season
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Re: Why am I getting contaminants? [Re: Sivarted]
    #22414824 - 10/21/15 05:58 PM (8 years, 3 months ago)

It can be anything. But if you want to eliminate all the possibilities do everything right. Make and sterilize your own jars. Eliminate that possibility. Inoculate in a proper still air box. I have no restriction issues in my box.. My armholes are 6 inches big, and could easily be bigger. In fact, having bigger armholes in a still room is recommended these days. The tote I use is a 110 qt tub. Don't use alcohol wipes at all. I haven't had any need to. All you need is a flame. I'd also use a torch over a lighter. Outside the box too!!

If you have a microscope you could see the cleanliness of the spores. You can also do agar. But that approach takes longer because you have to wait for things to germinate.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
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OfflineSivarted
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Re: Why am I getting contaminants? [Re: Mad Season]
    #22414867 - 10/21/15 06:08 PM (8 years, 3 months ago)

Yeah, definitely going to get a 110 tub soon.  I've been using mine with agar, and that was *ok* but not ideal.  Just did my first G2G though, and it was super awkward to do even 6 jars.  Not sure what the size is but it's smaller than the 66s I picked up for monos, so obviously it's tiny.

Lighter is a pain in the ass too.  I just picked up an alcohol burner and fuel, works great.

I get away without using my SAB because the bathroom is small, easy to wipe down everything and spray lysol and bleach everywhere.  It's like a giant SAB that I can walk into. :smile:


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InvisibleMad Season
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Re: Why am I getting contaminants? [Re: Sivarted]
    #22414881 - 10/21/15 06:12 PM (8 years, 3 months ago)

Yeah for cakes people did open air for the longest time.. And some still do. You couldn't do agar work in a bathroom like that tho. You might be able to get by with g2ging for awhile since grains aren't as prone to contams. But a still air box is as clean as it gets. You can do open air agar work in them ffs lol


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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InvisibleLocN9ne
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Re: Why am I getting contaminants? [Re: Mad Season]
    #22415140 - 10/21/15 07:02 PM (8 years, 3 months ago)

I just got another 110qt. From home depot for $2 because the bottom had a crack in it... fuck yeah


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InvisibleWilly Wonka
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Re: Why am I getting contaminants? [Re: LocN9ne]
    #22415741 - 10/21/15 09:16 PM (8 years, 3 months ago)

Quote:

LocN9ne said:
I just got another 110qt. From home depot for $2 because the bottom had a crack in it... fuck yeah





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AMU



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InvisiblePastywhyteMDiscord
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Re: Why am I getting contaminants? [Re: Willy Wonka]
    #22415887 - 10/21/15 09:49 PM (8 years, 3 months ago)

Quote:

Willy Wonka said:
Quote:

LocN9ne said:
I just got another 110qt. From home depot for $2 because the bottom had a crack in it... fuck yeah









Just caught his show the other week, took the wyfe too. Was her first time at a tech n9ne gig and damn it was killer.


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InvisibleWilly Wonka
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Re: Why am I getting contaminants? [Re: Pastywhyte]
    #22415993 - 10/21/15 10:21 PM (8 years, 3 months ago)

Pasty check your :pm:


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AMU



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Offlinespirit_shadow
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Registered: 08/15/11
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Re: Why am I getting contaminants? [Re: Mad Season]
    #22416006 - 10/21/15 10:25 PM (8 years, 3 months ago)

Quote:

Mad Season said:
Yeah for cakes people did open air for the longest time.. And some still do. You couldn't do agar work in a bathroom like that tho. You might be able to get by with g2ging for awhile since grains aren't as prone to contams. But a still air box is as clean as it gets. You can do open air agar work in them ffs lol



hey, I have done agar that way WITH success. no bullshit, yeah I had to be ocd and clean and tape off all air circulation spots(door, windows,vent) BUT it worked so :P


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ERROR 418 IM A TEAPOT.....(this account is automated, all posts related to illegal activities or advice thereof are strictly from numerous online sites and are for informational purposes only)- Circa 2011
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InvisibleMad Season
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Re: Why am I getting contaminants? [Re: spirit_shadow]
    #22416045 - 10/21/15 10:39 PM (8 years, 3 months ago)

Oh I see.. I just spray my box with water lol... Every time I talk about open air I direct people to this:
http://pharmlabs.unc.edu/labs/parenterals/techniques.htm

After reading all that it made me realize the world of molds is soo vast. Just saying that after knowing what all can be up in the air, I can't really ever trust open air again lol.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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InvisibleLocN9ne
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Re: Why am I getting contaminants? [Re: Pastywhyte]
    #22416053 - 10/21/15 10:42 PM (8 years, 3 months ago)

Quote:

Pastywhyte said:
Quote:

Willy Wonka said:
Quote:

LocN9ne said:
I just got another 110qt. From home depot for $2 because the bottom had a crack in it... fuck yeah









Just caught his show the other week, took the wyfe too. Was her first time at a tech n9ne gig and damn it was killer.



He puts on a pretty good show...very high energy...here goes my GPS info...sometime in the near future I am going to see slipknot and suicidal tendencies...I can barely contain my excitement... gonna be a hell of a show, got my nose broke at my last concert...


--------------------


Q&A
US vs. THEM

The more I learn, the less I know.


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Offlinespirit_shadow
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Registered: 08/15/11
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Re: Why am I getting contaminants? [Re: Mad Season]
    #22416089 - 10/21/15 10:58 PM (8 years, 3 months ago)

Quote:

Mad Season said:
Oh I see.. I just spray my box with water lol... Every time I talk about open air I direct people to this:
http://pharmlabs.unc.edu/labs/parenterals/techniques.htm

After reading all that it made me realize the world of molds is soo vast. Just saying that after knowing what all can be up in the air, I can't really ever trust open air again lol.



I feel ya man, im just saying its not impossible lol.


--------------------
ERROR 418 IM A TEAPOT.....(this account is automated, all posts related to illegal activities or advice thereof are strictly from numerous online sites and are for informational purposes only)- Circa 2011
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InvisibleWilly Wonka
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Re: Why am I getting contaminants? [Re: Mad Season]
    #22416181 - 10/21/15 11:29 PM (8 years, 3 months ago)

Quote:

Mad Season said:
Oh I see.. I just spray my box with water lol... Every time I talk about open air I direct people to this:
http://pharmlabs.unc.edu/labs/parenterals/techniques.htm

After reading all that it made me realize the world of molds is soo vast. Just saying that after knowing what all can be up in the air, I can't really ever trust open air again lol.Never again!!!




  A glove box is the shit!  A flow hood even better.  You  are right about that. Contams are everywhere!  We are trying to give the fungi (mycelium) the best fighting chance to live in the environment which we have created for it. 

Molds are everywhere.  Mushroom mycelium can fight of contminations in the wild.  If given a chance mycelium can fight off contmas in a lab for sure.  As mycologists we are trying to give the spores the best chance to live that we can.


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AMU



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OfflineEdmunter
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Re: Why am I getting contaminants? [Re: Willy Wonka]
    #22416496 - 10/22/15 02:25 AM (8 years, 3 months ago)

You should always do your work in a prepared SAB.  Flame the needle and wipe the injection port.

Im gunna just say it though, buy your self a pressure cooker and sterilise the jars your self.  It so easy and you can also use it to cook with.  Also even with a syringe its best to grow out a culture on agar to make sure and not waste your time.

Check out the videos in my link and read the contamination bit.


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OfflineDrCrumbs
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Registered: 10/25/11
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Re: Why am I getting contaminants? [Re: Edmunter]
    #22417312 - 10/22/15 09:13 AM (8 years, 3 months ago)

Can you post a pic?

I've done open air tons of times (clean room, allowed to settle down), way back when. Including syringes, making syringes, and agar work. I dont recommend it, I did have very good success. Not a huge deal when working with just syringes and ports though.

I never liked the oven tek. I feel like it stirs up the air more than anything.

I think the biggest problem with buying premade substrates/jars is the time from PC'ing till you get to knock them up.


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Offlinespirit_shadow
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Registered: 08/15/11
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Re: Why am I getting contaminants? [Re: Willy Wonka]
    #22417345 - 10/22/15 09:23 AM (8 years, 3 months ago)

Quote:

Willy Wonka said:
Quote:

Mad Season said:
Oh I see.. I just spray my box with water lol... Every time I talk about open air I direct people to this:
http://pharmlabs.unc.edu/labs/parenterals/techniques.htm

After reading all that it made me realize the world of molds is soo vast. Just saying that after knowing what all can be up in the air, I can't really ever trust open air again lol.Never again!!!




  A glove box is the shit!  A flow hood even better.  You  are right about that. Contams are everywhere!  We are trying to give the fungi (mycelium) the best fighting chance to live in the environment which we have created for it. 

Molds are everywhere.  Mushroom mycelium can fight of contminations in the wild.  If given a chance mycelium can fight off contmas in a lab for sure.  As mycologists we are trying to give the spores the best chance to live that we can.



I totally agree, but on the flip side I want my myc to be strong. strong enough to battle and consume a contam, which I have personally witnessed and ate the fruits of said myc :crazy2:


--------------------
ERROR 418 IM A TEAPOT.....(this account is automated, all posts related to illegal activities or advice thereof are strictly from numerous online sites and are for informational purposes only)- Circa 2011
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Offlinefiegtree
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Registered: 10/04/15
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Re: Why am I getting contaminants? [Re: spirit_shadow]
    #22417501 - 10/22/15 10:09 AM (8 years, 3 months ago)

Thanks for the replies everyone! I didn't expect to get this many at all. Does anyone have an answer as to how imperative it is to remove a suspicious jar from the chamber?


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If it can be destroyed by the truth, it deserves to be destroyed by the truth


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