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Offlinepsilohero
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Registered: 11/18/13
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Last seen: 9 years, 7 days
NAME THAT CONTAM!!!
    #19306180 - 12/21/13 02:45 AM (10 years, 2 months ago)

Ok so more specifically its "name the source of that contam". Every week only about 1/3 of my jars on average colonize. The rest of my jars develop what I assume to be a bacteria contam. They have a nasty dirty laundry raunchy wine type of smell with no real visible contam except they are slightly wetter under the needle holes. What would be the most likely areas (in order of likelyhood) that is the entry of this contam? Is this happening during steaming or during inoculation or for some more unknown reason?
A little info, I steam sterilize for 90 minutes and i used to do open air innocs but have recently started becoming more sterile and using a glove box in hopes to solve this problem. This has not improved my outcome thus far.
Any suggestions at all would be much appreciated as I have had this problem since I started (for about 8 weeks now) and am ready to throw in the towel because I have tried altering every variable I can think of.


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Offlineun-bear-able panda
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Re: NAME THAT CONTAM!!! [Re: psilohero]
    #19306270 - 12/21/13 03:32 AM (10 years, 2 months ago)

what tek are you using?
i recently switched from BRF cakes to popcorn and it's been a bit more successful from what i've noticed.


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Edited by un-bear-able panda (12/21/13 03:36 AM)


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Offlinepsilohero
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Re: NAME THAT CONTAM!!! [Re: un-bear-able panda]
    #19306340 - 12/21/13 04:01 AM (10 years, 2 months ago)

Im using pf/brf tek. I follow the guide with the proper updates that have been recognized on these forums. I dont want to have to run from my problems and switch to a different medium though, there should be a logical reason I am getting (what Im assuming to be) bacteria contam. Is there a way to tell if the problem is originating from the steam sterilization or inoculation? symptoms from either I could look for?


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OfflineKizzle
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Re: NAME THAT CONTAM!!! [Re: psilohero]
    #19307552 - 12/21/13 11:35 AM (10 years, 2 months ago)

How did you inoculate them? All from the same syringe? Flame the needle between jars? Needle didn't touch anything during the inoculation?


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Edited by Kizzle (12/21/13 11:42 AM)


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Offlineun-bear-able panda
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Re: NAME THAT CONTAM!!! [Re: psilohero]
    #19310080 - 12/22/13 12:18 AM (10 years, 2 months ago)

I didn't want to switch things up either but i had a similar issue and i am almost certain that the substrate i was using wasn't sufficiently sterilized with the steam, so i switched over to the popcorn still using steam to sterilize and it seems to have been working a lot better. If you want i can write up what i do but it's really not much different than any other popcorn tek found on the shroomery.


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Offlinepsilohero
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Re: NAME THAT CONTAM!!! [Re: un-bear-able panda]
    #19313981 - 12/22/13 10:28 PM (10 years, 2 months ago)

I have done some jars open air innoc and some in a still-air box but I havnt noticed that to make much difference.  I wash my hands with anti -bacterial soap and spray the area with oust.  Sometimes I flame the needle every 3 jars but once again not much change from when I dont flame the needle

Im starting the think I may not be getting them sterila from my stove.  I turn it on high for 15 mins, then turn it down to 3 for 60 mins originally but now 90 mins (didnt notice change in results). On 3 heat is provides a minor boil and steam.  The guide doesnt explain much about this except not to run the pot dry.  I was afraid of "over-heating" but now I dont think that is really an issue.  Thoughts?


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Offlineun-bear-able panda
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Re: NAME THAT CONTAM!!! [Re: psilohero]
    #19314304 - 12/23/13 12:25 AM (10 years, 2 months ago)

i do open air innoc in my kitchen after i wipe down the counter top with isopropyl alcohol and i like to use alcohol prep wipes on the syringe needles before i flame sterilize between each jar, it just makes me feel more confident that i didn't have any bacteria transfer from jar to jar. i also wear latex gloves that i wipe iso alcohol or hand sanitizer before i start handling the substrates and syringes.

that's exactly what i do with my stove, after the water is boiling i set it to 3 and set a stopwatch for 90 mins.

when i was using BRF not many of my jars would colonize and i started wondering what the issue could be. then i remembered one of those mushroom growing videos i watched told me that BRF contains lots of bacteria and that lots of contaminations come from the substrate itself, and that was the one thing i was too stubborn to change.

when i finally decided it was worth a try i bought some popcorn seeds and gave it a go. to my surprise, every single jar is colonizing awesomely after only 6 days. and i didnt have to change anything about my sterilization route which made me happy.


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Edited by un-bear-able panda (12/23/13 12:27 AM)


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OfflineKizzle
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Re: NAME THAT CONTAM!!! [Re: un-bear-able panda]
    #19315981 - 12/23/13 12:31 PM (10 years, 2 months ago)

If there are signs of bacteria focused around the inoculation points in particular and 0 sign of any kind of growth that's a pretty good indication it's being introduced during the inoculation. Still it's hard to say from what mentioned whether it'd be syringe or your inoculation technique. If you notice a correlation with a particular syringe contaminating all the jars it's inoculate, then it's probably that. Contaminated syringes are a common source of contamination.

As far as inoculation technique, you didn't mention wearing gloves which is definitely important as the human is the largest source bacteria in the house and bacteria covered skin is constantly flaking off into the air around them. Flaming between jars ensures if the needle becomes contaminated at some point during the inoculation you're only losing one jar because of it and not a bunch.

Another possible cause of bacteria is the dry vermiculite layer. If you make it with coarse vermiculite, don't make thick enough, leave an empty gap above it so chunks can easily move around, or if it gets damp during or after sterilization, those are all things that could lead to bacteria or other contaminants getting through it.

Make sure you're properly steaming them. The time starts when the pot is filled with steam. You need to use a lid that fits well to contain most of the steam.

Here's a recent version of the PF tek, there are a lot of version people have written with outdated and incorrect information you could look through it see if there's anything you may have missed.

Panda, if you're properly making and steaming your jars there's no reason you'd get a lower contamination rate switching popcorn which is far more difficult to sterilize. The whole point of PF style cakes is that you can get a 100% success rate from steaming them.


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Offlineun-bear-able panda
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Re: NAME THAT CONTAM!!! [Re: Kizzle]
    #19318147 - 12/23/13 08:03 PM (10 years, 2 months ago)

it's possible that i was making mistakes with the PF tek, but i personally dont think sterilizing the popcorn was any more difficult it was just more time consuming because of the soaking and boiling before filling the jars to steam. i'll get some new BRF and verm in the near future and see how it goes with some cambos using the link you provided to learn more about it. i've been using BRF for the past two years with limited success and i have high hopes for the next experiment now.


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Offlinepsilohero
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Re: NAME THAT CONTAM!!! [Re: un-bear-able panda]
    #19326631 - 12/25/13 07:28 PM (10 years, 1 month ago)

Im currently trying a control jar tek this time around.  I have tried a few jars at different heats.  Jars were left in the pot for 90 minutes to sterilize at heat settings of 3, 5 & 7, after having been brought to a boil for 15 minutes.  Im waiting a full week to open the jars but I believe I smell nastiness coming from control jars at each of the 3 settings.  Some weeks Ill have 40-50% of my jars colonize and some weeks Ill have 0% and I just dont really get it.  Whats even more weird is I used to grow years ago and I do everything the exact same but I had 95%+ success then..


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