|
Envypenis
Stranger

Registered: 10/29/22
Posts: 24
Last seen: 2 years, 1 month
|
70% iso doesn’t kill all bacteria
#28023689 - 10/29/22 09:32 PM (2 years, 2 months ago) |
|
|
Been experimenting with mycology for a little over a year now. It’s been a learning experience to say the least. This stuff makes growing world class weed look easy lol battled a lot already but my recent problem has to do with getting clean agar dishes. Had great success until a couple months ago.
I’ve improved on my pressure cooking technique. I do quart jars with new lids every time. Filled to within 1/2” headspace. I let steam blow out 10 mins to push out air of PC, then do a forty minute cook at 15 psi, wait for pressure to come down to 0 to remove. I get zero boil over, the lid is popped down and sealed before I even take the jar out of the PC. I have to assume my agar is safe as jams are done this way and will last for years on shelves.
I either wait for the agar to hit 130* or water bath until I’m ready. Have to bring it down to my tent, re sterilize everything and every time my hands leave that box for a second they get douched. Soak my lab Coat arms. I mean I can barely breathe in this tent with the alcohol fumes. Last batch i worked the stack of dishes from the bottom up. Still got half contaminated on the Amazon sterilized petri dishes. Had extra agar tried pouring into the jello Shots and got 60% clean - better than Petri dishes. Go figure..
I do all my sterile work inside a grow tent which I clean weekly with a bleach water mix. Stainless work table with air box from Amazon which has zippered clear vinyl sides, fabric bottom.. hepa 2500 sq ft filter inside the 4x8 tent. Positive pressurized - clean vented in with filtration 13 cloth material secured over air inlet. I’m always showered and no shoes in my tent. Clean clothes, lab coat with rubber bands over the cuffs with xtra long gloves up my wrists. I spray 70% iso all over the SAB. Soaking it to the point it’s dripping and the bottom is a puddle. I wipe and sanitize everything multiple times. I’ve tried brand new bagged and sealed x 2 manufactures disposable Jell-O shot cups as well as now the disposable but supposedly sterile PET petri dishes off Amazon. Still have contam all the way thru. Two batches ago 3 out of 25 jellos made it after a week sitting in my SAB.
Consistently the last three or four times I’ve poured I’m getting milky or cloudy spots within 24 hours on my agar. Some small raised shiny bacterial Spots as well. From what I’m reading in all the teks.. I’m way over killing it on sterilization.
Here is what I believe the issue is but I’d love to get some feedback and if anyone has experienced this…
70% iso is not killing all the bacteria in my SAB. According to these findings.. of 282 samples that were “disinfected” with 70% iso - 100 samples still contained microorganisms. Is it possible that I have an alcohol resistant bacteria in my SAB? Or am I losing my mind over something stupid that I’ve missed along the way here..
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4623738/
Edited by Envypenis (10/29/22 09:39 PM)
|
Brewtality



Registered: 12/06/20
Posts: 18
Loc: Your Neighbor's House
Last seen: 11 days, 6 hours
|
Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] 1
#28023705 - 10/29/22 09:42 PM (2 years, 2 months ago) |
|
|
Have you considered that it isn't a STERILE air box and instead a still air box. Are you turning the fan off when you go into the tent? Try slowing way down and trying not to disturb the air when working in the box. I have to remind my repeatedly to slow down or I my speed slowly increases. Have you tried making a video of your process. That might help you understand where things are going wrong.
|
Bajazly
Stranger... Than You Think



Registered: 09/02/22
Posts: 832
Loc: BC, Mexico
Last seen: 6 months, 27 days
|
Re: 70% iso doesn’t kill all bacteria [Re: Brewtality]
#28023711 - 10/29/22 09:46 PM (2 years, 2 months ago) |
|
|
Positive pressurized? You have a fan running thru your SAB? Filtered?
|
TheUsualSuspect
never tell me the odds


Registered: 09/11/22
Posts: 260
Last seen: 2 years, 5 days
|
Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
#28023721 - 10/29/22 09:51 PM (2 years, 2 months ago) |
|
|
Quote:
Envypenis said: Had great success until a couple months ago.
What changed from when you were getting success to now? Must have been a process or a piece of equipment.
Im not sure I completely undestand your whole process but are you using the grow tent as a still air box but also pumping air into it?
Quote:
Envypenis said: clean vented in with filtration 13 cloth material secured over air inlet"
that part isnt tracking well either.
|
hazyhorse
scoobin



Registered: 03/19/19
Posts: 3,822
Last seen: 2 months, 5 days
|
Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] 1
#28023753 - 10/29/22 10:08 PM (2 years, 2 months ago) |
|
|
you’re going way overboard dude. you can generally work in a dry, no prior cleaning SAB with bare hands & get perfect results. doesn’t hurt to be clean & wear gloves, but main thing is to work in a draft free room with no fans running. as someone else said, it’s a STILL air box, not STERILE air box. gravity is what keeps your media sterile, not clean air. you’d have to work in a lab grade HEPA filtered clean room wearing full PPE & shit for it to be sterile. you should not be gassing yourself out with chemical fumes man. you’re always going to have bacteria in your SAB because it’s not a sterile environment. as mentioned earlier, the air in the room being still & your motions being smooth, slow, & deliberate are going to make way more of a difference than bleaching out a grow tent & then running a fan.
-------------------- you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================
 
🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿
|
SingularFusion


Registered: 10/31/18
Posts: 1,303
Last seen: 1 month, 2 days
|
|
so you say your SAB has clear plastic and zippers?
Well I bet the walls move a little, this causes a suction and blowing type effect, sucking contam in through wherever there is negative pressure, which will be the arm holes in this case I guess. Why don't you build a simple SAB from a stiff plastic tub, I would bet your situation would improve
SAB works on still air... The concept of sterilizing it with iso is asinine, iso only gives things a good clean, sterile is not expected. Using the SAB successfully is all about having good technique and a still room around you. If you are blowing air in to your workspace(filtered or not), you don't have a still air box anymore and as such it works like you have seen it work, poorly
|
hazyhorse
scoobin



Registered: 03/19/19
Posts: 3,822
Last seen: 2 months, 5 days
|
|

i didn’t even realize it wasn’t a hard plastic SAB when i read that the first time. the flexible sheets are gonna stir up a ton of air even with subtle movements. that isn’t helping anything
-------------------- you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================
 
🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿
|
the man
still masked



Registered: 08/12/99
Posts: 6,778
Loc: C A N A D A
Last seen: 1 hour, 15 minutes
|
|
your issue is not the 70% rather the air movement. turn off the fan while working. is there soil/hydro in this tent aswell? imagine thats part of ur issue to as dont want cloth/carpet nevermind soil in the room.
the KEY is still air NOT cleaning the air to 100% sterile. u can rub ur hands with strik 9 and bleach and if wave them over agar they will contam 60% of the time.
|
TheUsualSuspect
never tell me the odds


Registered: 09/11/22
Posts: 260
Last seen: 2 years, 5 days
|
|
All that slopping cleaning chemicals everywhere and gassing yourself out, id stop doing that.
You seem willing to spend a little money on your hobby. If I were you I would get a small flow hood unit and put it inside that grow tent (and close off the outside air flow vents) and then you have a solid foundation to do work.
Get in the tent, zip it closed, let the flow hood run a bit and then start the lab work.
just my opinion
|
the man
still masked



Registered: 08/12/99
Posts: 6,778
Loc: C A N A D A
Last seen: 1 hour, 15 minutes
|
|
naa def dont need to close off vents with flow hood. u want air to move out and away easily not making eddies and swirling things up in a tiny room. anyhoo. SAB still air box NO moving air near ur box or made by fast movements. simple.
|
SirPsycho
Purple Belt in Google-Fu



Registered: 01/01/20
Posts: 7,340
Loc: Rent free in your head
Last seen: 2 days, 22 hours
|
Re: 70% iso doesn’t kill all bacteria [Re: the man]
#28023794 - 10/29/22 10:56 PM (2 years, 2 months ago) |
|
|
No shit it doesn't kill all bacteria. It's a sanitizer, it doesn't sterilize.
An SAB is a STILL AIR box. Not a clean air box.
--------------------
Ask about free prints: Ps tampanesis, subtropicalis, cubensis(ESS) and Pan cyanescens
Balance in life is like running on ice.
🅑🅞🅣🅣🅛🅔 🅖🅐🅝🅖
   
"Mist your balls and fan your asshole" - Pandaskis, 2023
|
Baba Yaga
Psychedelic Minion

Registered: 09/13/20
Posts: 4,490
Loc: Try 'n' Error Boot Camp
|
Re: 70% iso doesn’t kill all bacteria [Re: the man] 3
#28023803 - 10/29/22 11:09 PM (2 years, 2 months ago) |
|
|
You have this POS right?

This is the reason for your failure. You will be better off throwing this disgrace of SAB away. I can't believe those ass wipes are actually selling something like this and their advertising is making me throw up, "designed with you in mind", yeah right.
Sorry for the rant it's not your fault, but honestly get rid of this thing and build a SAB out of a rigid plastic tote. Reduce your alcohol use to a minimum and rely on good sterile technique......you know, the kind of technique you weren't able to achieve with this floppy SAB these dickheads sold to you.
|
matsc
Stranger



Registered: 09/17/12
Posts: 681
Loc: Arizona
Last seen: 1 year, 2 months
|
Re: 70% iso doesn’t kill all bacteria [Re: Baba Yaga]
#28023845 - 10/29/22 11:34 PM (2 years, 2 months ago) |
|
|
While I do agree with what folks are saying about the Positive-pressuretent and SAB, and that you are going a bit overboard at times....
My microbiologist heart requires me to say that, yes, there are in fact some bacteria that can survive 70% isopropanol exposure. Hell, when we buy Bacillus endospores to play with in my lab, they're literally sent as an alcohol suspension!
But they are a fairly niche group, and exposure time matters, and etc etc etc. In general, rubbing alcohol is fine as a surface sanitizer. Won't totally sterilize stuff, but useful tool in most situations.
And all that said, when doing lazy lab stuff at home, I just make pre-pour plates. Yeah the condensation in them is a little annoying, but when I only need a few plates at a time it's just easier for me. And I am VERY lazy most of the time
-------------------- My Trade List!
|
hazyhorse
scoobin



Registered: 03/19/19
Posts: 3,822
Last seen: 2 months, 5 days
|
Re: 70% iso doesn’t kill all bacteria [Re: matsc]
#28023850 - 10/29/22 11:38 PM (2 years, 2 months ago) |
|
|
that’s fine, but no one is saying there aren’t alcohol resistant bacteria, just that the problem you think you’re having & the problem you’re actually having aren’t related. even if the plates are pre poured, if you’re working in the conditions you described there’s still a huge chance for shit to be blown all over the open plate you’re working with. the clear plastic tote SAB with no drafts in the room is what you want
-------------------- you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================
 
🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿
|
matsc
Stranger



Registered: 09/17/12
Posts: 681
Loc: Arizona
Last seen: 1 year, 2 months
|
Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse] 1
#28023867 - 10/29/22 11:58 PM (2 years, 2 months ago) |
|
|
Quote:
hazyhorse said: that’s fine, but no one is saying there aren’t alcohol resistant bacteria, just that the problem you think you’re having & the problem you’re actually having aren’t related. even if the plates are pre poured, if you’re working in the conditions you described there’s still a huge chance for shit to be blown all over the open plate you’re working with. the clear plastic tote SAB with no drafts in the room is what you want 
Definitely agree with ya, but um... I'm not the original poster, just a mildly pedantic forum lurker I'm afraid! Sorry for the confusion
-------------------- My Trade List!
|
Tormato  
Chairman of the Bored




Registered: 07/01/17
Posts: 6,106
Last seen: 2 days, 11 hours
|
Re: 70% iso doesn’t kill all bacteria [Re: matsc] 1
#28023881 - 10/30/22 12:06 AM (2 years, 2 months ago) |
|
|
I'm just glad nobody mentioned peroxide  
-------------------- Helpful Threads
The Shroomery Store
Tormato's Q&A Thread Post Questions Here or PM me!
"Lately it occurs to me what a long, strange trip it's been." ~ Grateful Dead
Before you start...Do you have a Pressure Cooker and a Dehydrator? I highly recommend getting both!
|
Grind365
AlwaysGrinding

Registered: 08/07/22
Posts: 448
Last seen: 1 year, 11 months
|
Re: 70% iso doesn’t kill all bacteria [Re: Tormato]
#28023882 - 10/30/22 12:13 AM (2 years, 2 months ago) |
|
|
Peroxide
|
Tormato  
Chairman of the Bored




Registered: 07/01/17
Posts: 6,106
Last seen: 2 days, 11 hours
|
Re: 70% iso doesn’t kill all bacteria [Re: Grind365]
#28023890 - 10/30/22 12:27 AM (2 years, 2 months ago) |
|
|
Quote:
Grind365 said: Peroxide
You just had to didn't you. 

-------------------- Helpful Threads
The Shroomery Store
Tormato's Q&A Thread Post Questions Here or PM me!
"Lately it occurs to me what a long, strange trip it's been." ~ Grateful Dead
Before you start...Do you have a Pressure Cooker and a Dehydrator? I highly recommend getting both!
|
hazyhorse
scoobin



Registered: 03/19/19
Posts: 3,822
Last seen: 2 months, 5 days
|
Re: 70% iso doesn’t kill all bacteria [Re: matsc]
#28023897 - 10/30/22 12:41 AM (2 years, 2 months ago) |
|
|
Quote:
matsc said: Definitely agree with ya, but um... I'm not the original poster, just a mildly pedantic forum lurker I'm afraid! Sorry for the confusion 
LMAO ignore me, my bad friend!! my brain sometimes doesn’t register it’s a different person when people don’t have an avi. please continue your pedantry as you see fit
-------------------- you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================
 
🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿
|
Grind365
AlwaysGrinding

Registered: 08/07/22
Posts: 448
Last seen: 1 year, 11 months
|
Re: 70% iso doesn’t kill all bacteria [Re: Tormato]
#28023898 - 10/30/22 12:44 AM (2 years, 2 months ago) |
|
|
Quote:
Tormato said:
Quote:
Grind365 said: Peroxide
You just had to didn't you. 
 
Sorry. At least I didn't drop the link to the video of the guy touching petri dish after using iso, peroxide and just bare finger.
I may go find the video now though the world needs to know.
|
|