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InvisibleShamanBag
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I need help. Too much contamination too often.
    #26531257 - 03/12/20 03:22 PM (3 years, 10 months ago)

I really am looking for a system that has the least odds of contamination during the spawn/spawn to bulk phase.  My tubs more often than not contaminate with Trich so it must be a problem with my spawn.  I use a SAB and had good results a while back with MS syringes.  The tubs didn't contaminate but then I G2G'd and those jars, although they looked and smelled great, were apparently contaminated because the last 4 tubs I've done have all contaminated before the first flush.  I've got agar dishes going and I'd really prefer not to do the MS grows. 

I douse everything in 70 percent iso before I put them in the box and try to do everything I can to be as sterile as possible during the G2G or anything else I do in there but I can't take the tubs contaminating anymore.  I need to up my game so I can just get consistent good results. 

Do I need to get a laminar flow hood or build one?  Should I give up G2G and just use agar to inoculate?  What are the most consistent lowest chances of contamination techniques that I could employ?  I've followed teks but I'm ready to scrap the way I do things and start fresh with the best suggestions out there to minimize my chances of tub contamination.


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OfflineFrankRhizo
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Re: I need help. Too much contamination too often. [Re: ShamanBag]
    #26531271 - 03/12/20 03:29 PM (3 years, 10 months ago)

Yes agar will greatly increase your chances. You have to learn what a clean culture looks like and success will come.


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OfflineSockadin
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Re: I need help. Too much contamination too often. [Re: FrankRhizo]
    #26531300 - 03/12/20 03:44 PM (3 years, 10 months ago)

Try doing agar and 4 clean stransfers in go to grain. You can G2G agar wedge jars to expand to bulk.


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OfflineA.k.aM
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Re: I need help. Too much contamination too often. [Re: Sockadin]
    #26531375 - 03/12/20 04:33 PM (3 years, 10 months ago)

If the first jar was fine but then the four you g2g from it contamed it’s most likely happening during the grain transfer.


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OfflineMorbility
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Re: I need help. Too much contamination too often. [Re: A.k.a] * 1
    #26531511 - 03/12/20 06:31 PM (3 years, 10 months ago)

I second Sockadin's advice here: agar is definitely worth the minor effort and expense to get things clean and genetics narrowed down. It also opens up more options for putting myc to grain when the time comes. I'd start with looking at Bod's agar thread for an overview. I also preach the Josex Poke for dish transfers and going to LC for ease and simplicity.


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InvisibleLadysKnight
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Re: I need help. Too much contamination too often. [Re: Morbility]
    #26531592 - 03/12/20 07:26 PM (3 years, 10 months ago)

Be sure your plates are clean. Go agar to grain. Layoff the g2g.


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InvisibleShamanBag
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Re: I need help. Too much contamination too often. [Re: LadysKnight]
    #26532774 - 03/13/20 01:35 PM (3 years, 10 months ago)

These latest tubs are agar to grain and then G2G.  I am not sure it it's happening in the agar to grain or the G2G stage but I imagine if I elimimate the G2G stage then that is one more possible contamination point I could avoid.  I suppose I'll try just agar to grain and skip the G2G for a while.  I am hoping to get a setup and workflow that makes it as unlikely as I can to eliminate contaminated spawn and then tubs.  I've considered having a bucket of iso and just dipping the entire jars into it before transfer to the sab or just building a Laminar flow hood.  I eliminated oats and went back to WBS since every time I've used oats, and that is many, it's contaminated but I finally had a good couple tubs when I switched back to WBS.  I've also just switched to using plastic lids on my jars and so far they are easier to work with and I haven't had any issues with the Gas exchange aspect as the mycelium colonizes.


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OfflinecronicrFacebook
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Re: I need help. Too much contamination too often. [Re: ShamanBag]
    #26532817 - 03/13/20 02:00 PM (3 years, 10 months ago)

most contams I have seen from g2g are pretty blatant, those tricky ones can be a real bitch though but if it's consistent then u can usually safely say it's a culture issue, not going g2g won't save you if your culture is fucked.
when you work in a sab it's neither sterile or sanitary you work with physics so the sab just needs soapy water, wiping down your jars and dishes with iso is fine  but soapy water is what you need for the sab itself, iso drys too fast and water will trap contams and the soap will break the surface tension.
when I run into these issues I like switching to sterilized grows so I get bags and sterilize my substrates ect


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InvisibleDeckardCain
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Re: I need help. Too much contamination too often. [Re: cronicr]
    #26533141 - 03/13/20 05:24 PM (3 years, 10 months ago)

Maybe list which TEKs you’re following, I’m a noob here still, but I’ve never had trich in any tub. I ran like 4 g2g transfers deep off a ms syringe and while they seem to get more and more bacterial (my fruits grow really short and before the sub colonizes fully) but no problems with moulds until I leave my tub with a ton of aborts or side pins and they rot and eventually get a blue mould to them. Anyways my point I’m trying to make is maybe it’s your pasturization of your coir? Which tek do you follow, ive always used a Coleman lunch cooler preheated it first for 5-10 min then put the coir in with boiling water overnight stirring it up once.
Other than that I’d be looking around your work space for a mould colony, and going agar to see if that helps any.

Also I’ve heard wbs is one of the tougher grains to work with.


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