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Offlinethenilsmeister
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Fighting Contamination
    #18213634 - 05/04/13 02:41 PM (11 years, 17 days ago)

In the last month I have had atleast half of my jars contaminate, and am failing to find the source of the issue. Just to give you some info I prep rye via RR's tek and work in front of a 24x24 flowhood with 1000cfm blower dialed into laminar flow so the lighter bends at a 45 degree angle. I have tested it with incense and it seems to be pulling in some air from the sides but no matter how much I dial it down it still seems to have some pull, I dont even know if this is an issue or not because the smoke still does not enter the central workspace but is blown away as it reaches the filter. I let it run for an hour or more before working, lysol the room, spray the air with water, take a shower, use mouthwash, wear a mask, wear tyvek sleeves and wipe the whole area down with 90% alcohol.  I am working in a small carpeted apartment room but have no where else to work it is reserved for mycology and does not get heavy traffic but perhaps its just not possible to work in that space? My contaminates also line up with the species for example both my Oyster GM have gotten black pin mold 4 times in a row, my Hericiums have gotten trich two times in a row. Though the cultures appear to be clean perhaps some spores got deposited on them but did not have available agar to germinate? I have had pretty good success up until now, but this failure is seriously taking a tole on my mentality. Some more info is I PC for 2.5 hours for 9 quart jars, I always use agar-rye-sawdust-mycobag for edibles (though havnt gotten shit to fruit yet). Had to throw out my two oyster bags because they got what appeared to be wet bubble after looking very healthy but not pinning for 10 days. What the fuck am I doing wrong? I cant keep wasting materials and loosing my sanity.


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OfflineJeff
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Re: Fighting Contamination [Re: thenilsmeister]
    #18213674 - 05/04/13 02:50 PM (11 years, 17 days ago)

If your contams are culture specific I would suspect the cultures.
70% alcohol works better but I don't think that is your problem.
Depending on your filter you may want to up the flow to the point where it is not sucking back.  Higher flow, even if it blows the lighter out, is better than turbulent flow due to suck back IMO.


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OfflineeLeSDenes
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Re: Fighting Contamination [Re: thenilsmeister]
    #18233879 - 05/08/13 05:35 PM (11 years, 13 days ago)

You should leave an open agar plate in front of your flow hood for a minute or two then parafilm it and check in a few days if its contaminating.

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Offlinemycofever
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Re: Fighting Contamination [Re: eLeSDenes]
    #18234532 - 05/08/13 07:57 PM (11 years, 13 days ago)

I would just like to add that I do not think that it is your room. I use a room with carpet in it. Before you get started it is a good idea to shut off the air conditioner. Make sure that you let the air get still in the room. Once I sit down to work I wait a minute to let it get still again. I am still using a SAB. The only time that I get a contam is with wild prints Wild clones do not give me a problem. It may be your flow hood blowing contams all over your work. I hate hearing stuff like this it makes me wish that I was there so we could figure out what is going on. When I first got started I ran into a few contams. Now I a almost 100% success. You will get it. I suggest that you invest into some agar. You can get 1 pound of MEA from fp for like $75 with shiping that's what I buy it lasts a long time for the amount of work that I do.


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Offlinethenilsmeister
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Re: Fighting Contamination [Re: mycofever]
    #18235754 - 05/09/13 12:47 AM (11 years, 12 days ago)

Im toying around with things. I found a small vortex running along the edge of my flowhood which I seemed to be able to stop by placing my alcohol lamp there while I work. Tried ramping it up and choking it down but that didnt seem to stop it.

I use agar for everything btw. I hate LC's but might have to start working with them again if I cant seem to figure my issue out. Also, where do you live? I live in one of the wettest areas in the US so theres alot of mold spores here.

Thanks for the input people, I know that theres really no way for you to pinpoint my source but throwin ideas out there is helpful.


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Offlinethenilsmeister
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Re: Fighting Contamination [Re: eLeSDenes]
    #18235771 - 05/09/13 12:50 AM (11 years, 12 days ago)

Quote:

eLeSDenes said:
You should leave an open agar plate in front of your flow hood for a minute or two then parafilm it and check in a few days if its contaminating.




I left a plate open for a good 20 minutes but it was only in one location. It also got a couple bac dots but I think that was probably from me when I came to close it.


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Invisiblewildernessjunkie
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Re: Fighting Contamination [Re: thenilsmeister]
    #18235806 - 05/09/13 12:59 AM (11 years, 12 days ago)

Theres no shame in using an LC. As long as its a properly prepared and verified clean culture.

You have a flow hood though. With that said, I think your problem is with your cultures. Basic laminar flow should be enough to eliminate contaminants.

You might check the numbers and data on your flow hood with someone thats smarter than me though.

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OfflineeLeSDenes
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Re: Fighting Contamination [Re: wildernessjunkie]
    #18236720 - 05/09/13 07:07 AM (11 years, 12 days ago)

I live in a moldy house. The humidity is very high and we got
Moldy walls and furnitures. We clean them as much as we can but still theres loads of spores in the air and i have 100% success with g2g in front of a flow hood.
Maybe try to put plexi or glass to the sides and the top of the flow hood so the vortex on the edges can't pull air in. I have seen a few flood hood like that and they should work.

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OfflineRogerRabbitM
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Re: Fighting Contamination [Re: thenilsmeister]
    #18267703 - 05/15/13 07:25 AM (11 years, 6 days ago)

Quote:

lysol the room, spray the air with water, take a shower, use mouthwash, wear a mask, wear tyvek sleeves and wipe the whole area down with 90% alcohol




Switch to 70% alcohol as it works much better.

For bacteria try some antibiotic agar.

For molds, try making some agar at half normal nutrient strength.  This will encourage the mushroom mycelium to grow faster so you can grab a transfer sooner.  It usually takes just a few transfers to get a culture cleaned up.  You might also search for the 'hot agar tek' I've posted a few times.
RR


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Invisiblefastfred
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Re: Fighting Contamination [Re: RogerRabbit]
    #18267893 - 05/15/13 08:58 AM (11 years, 6 days ago)

Nice to see you back RR.  Any suggestions on the best sugar strength to use in your agar for the fastest growth?

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Offlinedrake89
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Re: Fighting Contamination [Re: fastfred]
    #18277907 - 05/17/13 09:08 AM (11 years, 4 days ago)

As people have said its not your room it's likely your cultures.  I laugh and cry and cringe when I hear about people who do the whole huge routine before working in front of the hood.  Mine used to be parked in the kitchen which is mold central.  Recently I've been doing work on my carport: outside!  All of my contam issues are either already in the culture dish, cause I was doing work in a terrible glove box previously, or they were on the outside of dishes, jars, slants, etc.  I have been reluctant to spray slants and dishes with alcohol before working with them because then my labeling will disappear but it's better than fighting contams. 

I've found that contams may be running with the mycellium of a couple of species so I may have to ditch them.  It's better than being frustrated.


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Offlinethenilsmeister
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Re: Fighting Contamination [Re: drake89]
    #18278315 - 05/17/13 11:07 AM (11 years, 4 days ago)

Ya I am finding that I have a couple hidden contams in cultures which I am trying to weed out. Having trouble getting the trich out of my only hericium culture though, might have to buy some antibiotic agar or try the sandwich or hot pour tek. Things are going much better now though, looks like I got 100% success on 16 qts of grain to grained King Oyster. Its been 5 days and they are all 90% or done :dancer:.

Does anyone know if its possible to prepare multiple batches of spawn bags or do they need to be nocced up as soon as the PC is opened?


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Offlinedrake89
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Re: Fighting Contamination [Re: thenilsmeister]
    #18278578 - 05/17/13 12:14 PM (11 years, 4 days ago)

I find it's ok to leave them out for a few days if they cool and seal in the steralizer.  Don't know about a PC.  Obviously spray them down before innoculating if they don't sit in front of a flow hood.


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