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Offlinemushpunx
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How do you keep your tyvek clean?
    #21773631 - 06/07/15 11:06 AM (8 years, 7 months ago)

I usually use tyvek sleeves with scrubs taken out of the dryer (used.to draw blood in a hospital so I have lots of scrubz laying around) but lately Ive been using those full tyvek suits with hoods that I got from a trade.

Usually I take my tyvek off and put into a gallon ziplock. The sleeves get cleaned alot because they go thru PC cycle in my spawm bags.

How do you guys clean them for work? Is a PC cycle the best option?


I get contams in during G2g despite my best efforts so just picking apart my process :shrug:


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InvisiblePastywhyteMDiscord
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Re: How do you keep your tyvek clean? [Re: mushpunx]
    #21773767 - 06/07/15 11:53 AM (8 years, 7 months ago)

Have you already verified that its not your culture? I don't run bags much but I do know the frustration of dealing with an unknown vector. Sometimes its not what you think at all. If you think its the G2G with the bags that is the point where it contams then I suggest the first thing you do is a grow with just agar to grain. If that works out consistantly then you can move on. But if any of those grows contam you might be looking at something else.


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Offlinemushpunx
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Re: How do you keep your tyvek clean? [Re: Pastywhyte]
    #21774069 - 06/07/15 01:25 PM (8 years, 7 months ago)

Well yea tell you the truth Im not 100% where its getting in at all, Ive just been trying to improve my technique in all areas to try to cover my bases.

Ive just started running bags so thats why Ive been kinda leaning towards it getting in during G2G, although I had my share of green mold when I was just working with jars.

This last run was really confusing because I was working with several different cultures. Kizzle sugguested that I run all my cultures through an antibiotic plate before I use them so Im going to give thay a shot too and see how it goes


Yea, I think I oughta give that a try and go from there. Like take my culture, grow it out on X amount of plates, then split each plate A2G into like 5 jars per tub.


As of late Ive been going wedge-pint master-4.5qt bag-tub


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InvisiblePastywhyteMDiscord
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Re: How do you keep your tyvek clean? [Re: mushpunx]
    #21774188 - 06/07/15 01:52 PM (8 years, 7 months ago)

I have suspected for a long time that the main source of mold before first flush is actually due to bacteria. I suspect that most people have bacterial spawn and don't even know it. Because it is possible to fruit through bacteria many people assume the spawn was clean but I have seen multiple flushes from bacterial spawn that never smelled off and fully colonized.


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Offlinemushpunx
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Re: How do you keep your tyvek clean? [Re: Pastywhyte]
    #21775038 - 06/07/15 04:53 PM (8 years, 7 months ago)

This is what Kizzle and I were talking about

But bacteria is usually easy to spot on a plate though yea? It must be hiding in the culture some how?

So yea I was gunna try mixing some gentamyacin into a sleeve of plates and putting my cultures thru a transfer or two on those before they go to grains, Im very curious to see how it goes

I wish I could video myself working and review it too see if Im making some silly error somewhere. I go through the works though, never cut corners.
Shroomery should be proud :lol:


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InvisiblebodhisattaMDiscordReddit
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Re: How do you keep your tyvek clean? [Re: Pastywhyte]
    #21775220 - 06/07/15 05:38 PM (8 years, 7 months ago)

Quote:

Pastywhyte said:
bacterial spawn that never smelled off and fully colonized.



:bigyesnod:
it exists.

I'm trying to get spore strip tests for autoclave validation, they're quite expensive and you need an incubator but I want to see just how long at which pressure it does take to get a grain jar sterilized. I feel like 90m at 15psi for more than a couple quart jars isn't enough.


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Re: How do you keep your tyvek clean? [Re: bodhisatta]
    #21775550 - 06/07/15 07:09 PM (8 years, 7 months ago)

If you're doing a no soak prep then it probably is bacteria. You're hydrating and then pcing so quick that the bacteria likely is just starting to soften and "germinate" by the time your culture is colonizing the grain it calls home and you're making a ticking bacteria bomb.

Bacteria becomes active, damages mycelium, mold spores germinate on damaged mycelia. Game over


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InvisibleMad Season
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Re: How do you keep your tyvek clean? [Re: maddchef]
    #21775613 - 06/07/15 07:26 PM (8 years, 7 months ago)

Not true. Just up the pc times to kill endospores. Endospores take more than a few days to germinate. Soaking it is just getting more bacteria in it during the soak causing the foul stench. If anything they'll make endospores during the pc cycle, so you have a higher chance of getting more endospores. We pasteurize to kill all molds but bacteria. Why do we pc for 90 minutes (i upped to 2 hours with great success) when a 30 minute boil at atmospheric pressure kills all molds? To kill the endospores.

90 minute pc:
120 minutes:

No soak method.


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Edited by Mad Season (06/07/15 07:28 PM)


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InvisiblebodhisattaMDiscordReddit
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Re: How do you keep your tyvek clean? [Re: Mad Season]
    #21775664 - 06/07/15 07:37 PM (8 years, 7 months ago)

endospores can germinate in a range of times from minutes to hours.


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InvisibleMad Season
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Re: How do you keep your tyvek clean? [Re: bodhisatta]
    #21775684 - 06/07/15 07:40 PM (8 years, 7 months ago)

Well I'm pretty sure it was you bod who said they don't germinate, or wouldn't germinate enough during a 24 hour soak to make a difference. Just to clear things up


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Edited by Mad Season (06/07/15 07:44 PM)


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InvisiblebodhisattaMDiscordReddit
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Re: How do you keep your tyvek clean? [Re: Mad Season]
    #21775707 - 06/07/15 07:46 PM (8 years, 7 months ago)

it's not all or nothing. some endospores will germinate quickly some will take hours. just like with spores. spores don't take a predefined time to germinate all at once.

most of the time they don't just up and germinate, but in optimal conditions it could be minutes. Could I tell you what those splits percentages etc.. are no, but there's plenty of evidence out there about endospore germination and it only takes days if conditions are not met. when lowest suitable conditions for germination are met some may be more fast than others when very suitable conditions for germination are met lots will germinate in minutes to hours some will be quicker and some slower still too.


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InvisibleMad Season
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Re: How do you keep your tyvek clean? [Re: bodhisatta]
    #21775723 - 06/07/15 07:49 PM (8 years, 7 months ago)

Sounds good :smile: gotcha. And it basically just keeps making more bacteria and as a natural defense it'll keep endospore population up. Altho not as much during harsh conditions, they're still there. Meaning heat in the end is how you get rid of them.


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Offlinemushpunx
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Re: How do you keep your tyvek clean? [Re: Mad Season]
    #21775754 - 06/07/15 07:58 PM (8 years, 7 months ago)

Are we talking about bacteria hiding out in our agar cultures?


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Re: How do you keep your tyvek clean? [Re: Mad Season]
    #21775769 - 06/07/15 08:01 PM (8 years, 7 months ago)

autoclave testing uses vials with very heat tolerant endospores that are non pathogenic. you put them in during a cycle and aferwards you incubate those spores on agar made specifically for their germination usually you incubate at something like 133F as that's the temperatures those particular stearothermophilus endospores germinate at. there's actually some fungi that have spores that need heat treatment to germinate like byssochlamys which infects pasteurized juice as the pasteurization process activates the spores.

so anyway if you're worried about sterilization and endospores, they test autoclaves with the hardest to kill endospores.


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Re: How do you keep your tyvek clean? [Re: Mad Season]
    #21775829 - 06/07/15 08:12 PM (8 years, 7 months ago)

Honestly, I don't believe in wearing tyvek, and in fact, as unpleasant as the visualization may be, I do all my G2G and even cloning work in the buff, or at most with a clean pair of tighty whities, lol. Well that and a hair net and a freshly washed/bleached bandana covering my mouth, look kinda like a masked streaker, lol.  I also work in open air, YES that's right, no glove box and DEFINITELY no laminar fucking flow hood either (what an immense waste of money, imo).

The main goal is to create an environment where there is ZERO air movement.  To this end I have a small bare room, no carpet, all vents have been sealed, and the window is also completely sealed with tape. Also have weather-sealed around the door to the room and have one of those tubular seal things that slides under the door so that it's completely air-tight for all intensive purposes.  Also make sure the AC/heating is turned off to minimize air movement within the house.

Then, about an hour before I start the process, I make sure all jars and work surfaces are wiped down with iso alky, and then I'll use about a 1/2 can of Lysol disenfectant to spray the entire room (and the bathroom too).  I then leave and take a shower, use a lufa to scrub most of my upper body (great for your complexion, btw) towel dry with a freshly washed/bleached towel, put on slippers, a hair net and surgical gloves then go directly to my clean room.  Open and close the door slowly, and then put on the sterilized bandana and tightly cover my mouth.

Sound like a lot of work?  Well yes, somewhat, although it's not that bad and the trade off is that IT IS FAR AND AWAY THE EASIEST WAY TO ACTUALLY DO THE WORK, MUCH EASIER IN MY XP THEN WORKING WITH GLOVE/SHMUV BOX.  So much easier to move around and see what you're doing as well.  And whether you believe it or not, I've done countless G2G transfers as well as cloning and even some agar work this way, despite "experts" opinons to the contrary.

And sorry if this is a bit of a hi-jack, but to the OP's original question, you don't need a tyvek suit.  Wipe down/wash your arms and hands with iso-alky, use latex gloves, also wiped with alky, and when doing G2G, wipe your spoon/utensile with alky between transfers and you should have no problem bringing contams into your workspace.


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InvisiblebodhisattaMDiscordReddit
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Re: How do you keep your tyvek clean? [Re: fearnoevil]
    #21775838 - 06/07/15 08:14 PM (8 years, 7 months ago)

never seen a naked scientist before but I'll let them know your theories. most of us have 99% success rates and we don't get all buffalo bill when doing culture work.


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Re: How do you keep your tyvek clean? [Re: Pastywhyte]
    #21775924 - 06/07/15 08:33 PM (8 years, 7 months ago)

Quote:

Pastywhyte said:
but I do know the frustration of dealing with an unknown vector. Sometimes its not what you think at all..



this.  one time, i was having contamination problems for a few weeks.  i figured out it was because i was trying to be cheap and re-use my gloves.  :facepalm: even though i was using alcohol to clean them, i would still get contams.  probably because the micro-tears in the latex caught bacteria.    it literally could be anything.  and i doubt it's your tyvek sleeves.


btw mushpunx, making a video is a great idea.  you should do it!  i bet we could all learn a lot from watching it and critiquing it. do eeeeeiiiit.


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InvisiblePastywhyteMDiscord
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Re: How do you keep your tyvek clean? [Re: bodhisatta]
    #21775998 - 06/07/15 08:47 PM (8 years, 7 months ago)

Quote:

bodhisatta said:
Most of us have 99% success rates and we don't get all buffalo bill when doing culture work.




We don't?  :derfase:


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Offlinemushpunx
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Re: How do you keep your tyvek clean? [Re: Pastywhyte]
    #21776164 - 06/07/15 09:22 PM (8 years, 7 months ago)

Well the reason Ive been using tyvek suit as of late is because (besides the fact I was given a bunch) it covers you up so well that, if the suit is clean you dont have to shower/ don freshly washed clothes when you work.
But of course that only works if the suit is clean haha.

Like I said I normally shower and wear freshly washed scrubs, mask, gloves, hair net and tyvek sleeves.

I do my work in front of a flow hood. If its just agar plates or even agar to grain innoculations Im working on it isnt the biggest deal, but when it comes to G2G (especially into spawn bags) its easier for any nastys that happen to be on my sleeves to get blown around amd get into my work. 

So yea, if Im scrubbing up and going through the works, but if my sleeves arent clean I could see it being a serious vector yea?


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Re: How do you keep your tyvek clean? [Re: mushpunx]
    #21776198 - 06/07/15 09:28 PM (8 years, 7 months ago)

Procedure is more important. Don't matter how well you showered or laundered shit, if your stirring up air currents or moving incorrectly in front of the hood its gonna be a problem.

Clean tyvek sleeves in your bags that then get sterilized ain't that important. I don't use bags much but again, any contams on them are being killed. Now if the pores opened up too much while PCing maybe that's the issue but, even then I don't think its likely.

You seemed less than confident that your masters were truly clean. I would look there. Sounds to me like a bacteria issue, especially if your really loading a lot of bags in at once or using grain with a massive endospore count. Might need to really up them PC times. For me, even a longer PC cycle didn't solve my endospore issues with a certain batch of grain, I needed to get really creative.


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