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InvisibleJessicaRabbit88
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Help me understand HEPA filters and best SAB sterile work area setup
    #26998184 - 10/22/20 03:39 PM (3 years, 3 months ago)

Before I moved and over the course of 4 months, I successfully produced 500+ grain jars with 4 contaminated jars (during my first month of this hobby). Now 40% of my jars and 25% of my agar work gets contaminated.

This is my new setup:



I pull the plastic curtain shut while I work —this is something I didn’t have before my move.
Is this a problem?

Each apt unit has its own separate HVAC system and as you can see, the filter is right above my SAB area. The air gets sucked up through that vent and pushed out thru vents in various parts of my apt. I turn off the air for a good hour before I do any sterile work.

The filter is NOT A HEPA filter —SHOULD I BUY AN HVAC filter ($110 on amazon)?

Or move my SAB?


Windows are tight and I work from home, live alone, no pets.

I’m a bit of a clean-neat freak and practice good sterile techniques:

- vacuum
- scrub down my area
- run a small HEPA air purifier for an hour
- shower, brush teeth (sometimes even wear a mask)
- wear new sterile latex gloves (Costco 400 pairs)
- continuously wet gloved hands with 70% ISO when I move hands outside of SAB
- wipe down jars with iso before and after putting them in SAB
- use sterile blades (packaged individually)
- spray entire SAB with soapy water
- use elevated cookie rack and small glass jars to elevate petris and Pasty plates
- never allow gloved hands to go over open jars or petris
- PC @ 17-18 psi for the recommended times (no shortcuts)

Jars, tubs and petris are kept in a good size walk-in closet with nothing else.

Sucks I have carpeting but I had carpeting before with no issues.


Edited by JessicaRabbit88 (10/24/20 09:57 AM)


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Offlineedgar1337
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: JessicaRabbit88]
    #26998279 - 10/22/20 04:31 PM (3 years, 3 months ago)

So you moved locations and suddenly got an influx of contamination? Sounds like there might be something in your place. Especially if what you say is true; as a noob you only had a few contaminations out of 500 jars before moving. If your in an apartment that might be why, a lot of apartments are filthy and don't have proper upkeep. Some tenants are total pigs, and tend to only do surface, aesthetic cleaning and not proper sterilizing before moving out.

I've been in apartments that had pipe issues that led to mold in the walls, and the entire block was displaced for a couple days. Considering your sudden influx of contaminations, if you are using the same teks, same practices, same grains, but are in a new place, I'd consider the agitator to be the latter.

But I'm also a noob myself, and not qualified to say.


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

"Mistakes are, after all, the foundations of truth, and if a man does not know what a thing is, it is at least an increase in knowledge if he knows what it is not."

-Carl Jung


"The current state of knowledge is a moment in history, changing just as rapidly as the state of knowledge in the past has ever changed and, in many instances, more rapidly."
-Jean Piaget


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InvisibleJessicaRabbit88
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: edgar1337]
    #26998540 - 10/22/20 07:19 PM (3 years, 3 months ago)

I was in an apartment before and had excellent results.

Then I moved to an old, house with hardwood floors which I steam cleaned daily. The house had been gutted to the studs but the area was very windy @nd dusty @nd I started to have some contam issues with the 2nd flushes of my tubs.

I moved 2 months later and into this apt and it’s more upscale, in a gated community and newer than either places I lived before.

I noticed the oat grains I was buying are so dirty and contain a lot of crap. Also, I did just start to use millet and the first batch of jars I made colonized fine. Now even G2G gets 40% contaminated.

Is my SAB work area in an ideal spot or should I move it?

Would installing a HEPA filter help?

I’ve read many responses by TCs insisting it’s not your environment but rather your sterile technique.


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Offlineshroomrookie
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: JessicaRabbit88]
    #26998645 - 10/22/20 08:36 PM (3 years, 3 months ago)



IMO any type of open air inlet above the SAB is bad. Dust and bad spores collects at the entrance to the vent. This is the problem.


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


Edited by shroomrookie (10/22/20 08:38 PM)


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Offlineedgar1337
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: JessicaRabbit88]
    #26998897 - 10/22/20 11:54 PM (3 years, 3 months ago)

Quote:

JessicaRabbit88 said:
I was in an apartment before and had excellent results.

Then I moved to an old, house with hardwood floors which I steam cleaned daily. The house had been gutted to the studs but the area was very windy @nd dusty @nd I started to have some contam issues with the 2nd flushes of my tubs.

I moved 2 months later and into this apt and it’s more upscale, in a gated community and newer than either places I lived before.

I noticed the oat grains I was buying are so dirty and contain a lot of crap. Also, I did just start to use millet and the first batch of jars I made colonized fine. Now even G2G gets 40% contaminated.

Is my SAB work area in an ideal spot or should I move it?

Would installing a HEPA filter help?

I’ve read many responses by TCs insisting it’s not your environment but rather your sterile technique.



So it might not be the location then. Dirty grains shouldn't matter if you are prepping and sterilizing them properly. I don't think a whole lot of bacteria can survive 12-24 hour soaks/boiling water/120min in a 200 degree pressure cooker.

In my limited experience, flowhoods are not necessary. Just wiping and spraying the workspace and air with 70% ISO a few times and turning off all fans works. The vectors for contamination seem to mostly come from pre-inoculation methodology. Inoculating is the briefest part of the whole thing, so it naturally has the smallest window for contamination.

Forget about a make-shift flowhood, get your still air box away from dirty vents or AC's, put it on a large flat surface (like a table), wipe the table with ISO, then spray the air and the table with ISO and leave it there. Wipe down your SAB, then spray it with ISO and leave the droplets. If you properly sterilized a viable medium (like bird seed/rye/whatever your using), it is unlikely to get contaminated in the 5 seconds it takes to move the jars from your pressure cooker to the SAB.

Inside the SAB the main vector for contamination is your hands and arms. Wearing latex gloves is ideal because you can be spraying your hands with ISO constantly and letting it evap in the SAB, and it won't irritate your skin. This advice is no doubt redundant, as you already know it all if you've done so many successful jars. But it can be easy to become complacent or lazy with sterile technique.

You mentioned local grains seeming crappy in quality - so it sounds like the grains you are using are different now. That could also be the cause of the influx in contams. Try and get the same brand that worked for you before. Or find them online.


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

"Mistakes are, after all, the foundations of truth, and if a man does not know what a thing is, it is at least an increase in knowledge if he knows what it is not."

-Carl Jung


"The current state of knowledge is a moment in history, changing just as rapidly as the state of knowledge in the past has ever changed and, in many instances, more rapidly."
-Jean Piaget


Edited by edgar1337 (10/22/20 11:58 PM)


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OfflineBloo
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: edgar1337]
    #26998920 - 10/23/20 12:07 AM (3 years, 3 months ago)

When do they get contaminated, within 2-5 days, or after a week or more?

Do containers that are never opened for inoculation contaminate as well?

Is the contamination the same in most of the containers- does one type predominate, or is it variable, sometimes with more than one organism?

Is the contaminant mostly found in the middle, on the bottom, on the top?

What happens if you place inoculated jars in a clear plastic bag immediately after inoculation?


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OfflineForresterM
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: shroomrookie]
    #26999026 - 10/23/20 02:29 AM (3 years, 3 months ago)

Quote:

shroomrookie said:


IMO any type of open air inlet above the SAB is bad. Dust and bad spores collects at the entrance to the vent. This is the problem.




:whathesaid:

Although it's not so much the presence of dust and spores on the air vent, it's the fact that it's moving all the air around, right above your STILL AIR box. 

Might not be what's causing your problems, but it's a good place to start.


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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InvisibleJessicaRabbit88
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: edgar1337]
    #26999100 - 10/23/20 04:50 AM (3 years, 3 months ago)

Thank you all for your input.

EDGAR —the brand of oats is exactly the same... “something Pride”... I read other comments that this brand’s quality has gone downhill and to find “race horse quality” oats. That’s next on my list.

II definitely clean all surfaces and wipe my SAB down and spray with soapy water. Then spray 70% ISO inside. All fans are off and I saturate my latex gloves with ISO as well as paper towels I use inside my SAB and wipe the jar threads as I unscrew the lids. I make sure not to let my hands go over open jars or petris.

I sterilize millet for 90min @ 17-18 psi. I’ve been using Fooman’s grain prep with a 30min soak vs 20min since millet is such a hard grain. It’s been coming out pretty good. Plump but not overly hydrated.


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InvisibleJessicaRabbit88
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: JessicaRabbit88]
    #26999107 - 10/23/20 05:05 AM (3 years, 3 months ago)

BLOO — they get contaminated after 8 to 14 days. Sometimes they stall, so I’ll shake and I’ll see dreaded grey-black grain. Sometimes green and other times orange. Or the bottom will look gooey. Or the oats will look gooey after a shake.

I seem to be having a little more success on my current 120 jars. Lost about 24. I attribute it to dialing in the right hydration for millet. As for the oats, I’m letting them dry out for 2 days in the fridge (after letting them drain). I’ve been using Bod’s oat tek and it had been working perfectly. I posted pictures when I first started and he said my grains looked like they should.

I have not placed the inoculated jars in an clear plastic bag... Won’t they suffocate?


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InvisibleJessicaRabbit88
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: JessicaRabbit88]
    #26999128 - 10/23/20 05:37 AM (3 years, 3 months ago)

SHROOMROOKIE & FORRESTER — I haven’t felt comfortable with that vent from the start. That vent pulls the air in and through a filter (not HEPA) and why I asked if it would be a good investment to buy one. But when I’m working, the fan is off and the air is still...

I shall go ahead and setup my SAB in a different room, away from all vents. I’m pretty anal when it comes to cleanliness and following procedures. I feel that’s why I’ve had the success I’ve had.

Once I get these contam issues resolved, I plan on switching to bags and continuous vet syringe. Since I don’t have a flow hood, I’ve been sealing my bags prior to PCing and they’ve been coming out great. The grains actually look better in the bags than in the jars. More evenly hydrated, not sticky or clumpy whatsoever.

But sucking up LC in a 60ml syringe was nerve wracking! Plunger was so hard to pull up. I ended up tilting the jar. The second time I tried, I used a lid with 2 layers of microspore tape, flame sterilized the needle until it was red hot, gave it a few seconds to cool and punctured through the tape, kept the jar upright and inoculated a couple of bags (covered the hole with packing tape (as per a tek I read on the shroomery)... I’ll update you in a few days.

Boy I love this community! You guys have all been great!


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OfflineForresterM
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: JessicaRabbit88]
    #27000230 - 10/23/20 06:34 PM (3 years, 3 months ago)

Yeah LC work can be nerve wracking after you read all the scares of using it :smile: 

I'm a big LC fan though, and grain in bags is awesome if you can get it to work.  Good luck with the SAB issues I'm sure you'll figure it out.


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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Invisibleeatyualive
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: Forrester]
    #27000925 - 10/24/20 08:10 AM (3 years, 3 months ago)

Make sure you turn off hvac prior to doing work. Cant be a worse spot to put that SAB with the vent right over it. If its off, no issue. You dont need plastic curtains. I dont think it will cause any issue but definitely not needed. I use a fold out table and a 110 quart sab. Then pack it all up when im done.
You dont need to run an air filter prior to the sab session. It cant hurt hut as long as the air is still is what matters. All i do is turn off the air. I shower and wash my arms up to my elbows with hot water and soap. I also use gloves. Not everyone does but i see those people having a far higher contamination rate than what i experience. I also just douse the ceiling, walls and floor of sab with 70 iso. I put cookie racks in to elevate the work area, although it isnt necessary.

https://www.shroomery.org/forums/showflat.php/Number/26488885


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Edited by eatyualive (10/24/20 08:16 AM)


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InvisiblebodhisattaMDiscordReddit
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Re: Help me understand HEPA filters and best SAB sterile work area setup [Re: JessicaRabbit88]
    #27000958 - 10/24/20 08:34 AM (3 years, 3 months ago)

Do you find yourself holding your breath while doing SAB work?
Stop it. Will immediately make a big difference


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