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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Sterile Equilibrium Returned To The Lab!
#21910837 - 07/07/15 05:44 PM (8 years, 6 months ago) |
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When several breakouts occur with the mold. It's time for some "In with the new & out with the old". A lot of time, and some bleach.. and low & behold Sterile equilibrium returned for clean cultures to unfold.

Every corner, crevasse, nook, cranny, shelf, surface, tool, and media vessel wiped with an alcohol soaked paper towel. All 6 surfaces of the room were wiped with a strong bleach solution using a white wash cloth solution TWICE! The flowhood was ran a hour before entry into the lab. Flowhood was kept running during the cleaning, and was kept running for up to a hour after the cleanse was finished.
Edited by hamloaf (07/08/15 06:30 PM)
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TravelAgency
The ongoing "wow"

Registered: 12/25/10
Posts: 4,431
Last seen: 11 months, 23 days
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Re: Sterile Equilibrium Returned To The Lab! [Re: hamloaf]
#21910855 - 07/07/15 05:47 PM (8 years, 6 months ago) |
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Beautiful! When I move into a larger place I'm totally doing that!
What type of procedures do you use for personal cleanliness when entering your room? Go all Walter White or more of a general shit shower shave scenario?
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Re: Sterile Equilibrium Returned To The Lab! [Re: TravelAgency]
#21911274 - 07/07/15 07:12 PM (8 years, 6 months ago) |
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Thanks, TravelAgency. Procedures used for personal cleanliness taken before lab is entered are flowhood is ran for 2 hours, shower, brush teeth (in shower), then freshly laundered clothes are worn (scrubs). A two gallon "dish tub" is filled with warm soapy water pre getting into the shower, and is placed at entrance of lab. Foot bath's purpose is for rinsing feet JUST before entering lab. Once lab is entered hands are gloved, tyvek wrist sleeves cover arms, and surgical face mask is put on. Ready to go to work! Before any tools, and/or media vessels are placed into the direct downstream of the flowhood they are wiped thoroughly with an alcohol soaked paper towel.
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TravelAgency
The ongoing "wow"

Registered: 12/25/10
Posts: 4,431
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Re: Sterile Equilibrium Returned To The Lab! [Re: hamloaf]
#21911397 - 07/07/15 07:35 PM (8 years, 6 months ago) |
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Awesome! Thanks for the info! I've read a lot of your write ups ham- thanks for all the help you provided to myself and I'm sure a litany of others.
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Re: Sterile Equilibrium Returned To The Lab! [Re: TravelAgency]
#21915433 - 07/08/15 03:04 PM (8 years, 6 months ago) |
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Yea, man. No problemo. A couple of things that were forgotten to be mentioned. As measures to maintenance of the lab's sterile equilibrium the flowhood is set to a timer to come on for two hours a day. Once at noon for an hour, then once at midnight for an hour. Also, all surfaces of lab including shelf tops are wiped once a week thoroughly with an alcohol soaked paper towel.
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Sterile Equilibrium Returned To The Lab! [Re: hamloaf]
#21916174 - 07/08/15 06:02 PM (8 years, 6 months ago) |
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Quote:
hamloaf said: Thanks, TravelAgency. Procedures used for personal cleanliness taken before lab is entered are flowhood is ran for 2 hours, shower, brush teeth (in shower), then freshly laundered clothes are worn (scrubs). A two gallon "dish tub" is filled with warm soapy water pre getting into the shower, and is placed at entrance of lab. Foot bath's purpose is for rinsing feet JUST before entering lab. Once lab is entered hands are gloved, tyvek wrist sleeves cover arms, and surgical face mask is put on. Ready to go to work! Before any tools, and/or media vessels are placed into the direct downstream of the flowhood they are wiped thoroughly with an alcohol soaked paper towel.
sounds like my clean work ritual.
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: Sterile Equilibrium Returned To The Lab! [Re: eatyualive]
#21916240 - 07/08/15 06:21 PM (8 years, 6 months ago) |
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Very nice ham! Can't wait until I have an actual lab and not just a spare bedroom full of cultivation equipment
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Let it grow! Let it grow! Greatly yield! What shall we say, shall we call it by a name As well to count the angels dancing on a pin Water bright as the sky from which it came And the name is on the earth that takes it in DOG FOOD AGAR MY ELECTRIC INOCULATION LOOP
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Sterile Equilibrium Returned To The Lab! [Re: eatyualive]
#21916251 - 07/08/15 06:25 PM (8 years, 6 months ago) |
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weak bleach is sometimes more effective at killing, say you buy bleach that's 4-6% 6% being clorox's strength. if you take clorox and do a 1:10 dilution it's more effective than it is mixed 1:1 with water.
bleach has a PH of around 11-12 but if you acidify it (this is what confuses people here, hang on) it becomes much stronger at killing.
hypoclorite in tap water around ph7 is much more active as the dissociated ions and available clorine than in a solution 1:1 with water which has very high pH still. 1:4 works well, bleach is stored at full strength or even 1:4 to last longer, lasting longer because it's not dissociating and chlorinating as much, the dissociation and free chlorine do the disinfection and water acts as the acidifying agent relative to the pH of bleach at bottle strength.
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Sterile Equilibrium Returned To The Lab! [Re: bodhisatta]
#21916291 - 07/08/15 06:35 PM (8 years, 6 months ago) |
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i use a 2-3% bleach solution in a spray bottle laying around handy. bleach wipes are also handy and easy.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Sterile Equilibrium Returned To The Lab! [Re: hamloaf]
#21916299 - 07/08/15 06:38 PM (8 years, 6 months ago) |
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Quote:
hamloaf said: Yea, man. No problemo. A couple of things that were forgotten to be mentioned. As measures to maintenance of the lab's sterile equilibrium the flowhood is set to a timer to come on for two hours a day. Once at noon for an hour, then once at midnight for an hour. Also, all surfaces of lab including shelf tops are wiped once a week thoroughly with an alcohol soaked paper towel.
have you seen the micro droplet disinfectant technology yet? you can peroxide fog a room to dramatically reduce contam loads and get near sterile environments but you don't want to accidentally lolercaust yourself
as long as the pH of your bleach solution is at or near 7 it will be quite effective for immediate use.
Edited by Trusted cuItivator (07/08/15 06:38 PM)
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: Sterile Equilibrium Returned To The Lab! [Re: eatyualive]
#21916300 - 07/08/15 06:38 PM (8 years, 6 months ago) |
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Lysol wipes are the best thing ever for sterile work. I even use fresh lysol wipes as my work surface inside the SAB. I used to use baking racks but they're clumsy. Now I just use the baking racks for making slants.
--------------------
Let it grow! Let it grow! Greatly yield! What shall we say, shall we call it by a name As well to count the angels dancing on a pin Water bright as the sky from which it came And the name is on the earth that takes it in DOG FOOD AGAR MY ELECTRIC INOCULATION LOOP
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Re: Sterile Equilibrium Returned To The Lab! [Re: bodhisatta]
#21918600 - 07/09/15 09:27 AM (8 years, 6 months ago) |
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That's good information over the desired concentration of bleach in order to most effectively kill invader micro-organisms, and their spores. Thanks for dropping that bit of knowledge in this thread, bod.
It's understood, and completely followed what you are saying about dropping the ph of bleach by acidifying it to make it more effective. What was meant when I said a "strong bleach solution" is more like a 1:5-7 bleach to water ratio. What you said will be kept in mind next time bleach solutions are mixed with the intents to use the solution as a lab disinfectant. Thanks.
The anti-contam fogger, and chemicals have been heard of, yes. Some of the guys (fellow Tribesmen) use a few of those chemicals and have foggers set up in their fruiting areas. In this cultivator's unique situation though there isn't enough room in the lab to fit such a device, so until larger real estate is had where a larger lab can be constructed I have to go with manually cleaning wiping down surfaces and running the flowhood in the previously described manner in order to keep the lab's sterile equilibrium maintained.
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TravelAgency
The ongoing "wow"

Registered: 12/25/10
Posts: 4,431
Last seen: 11 months, 23 days
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Re: Sterile Equilibrium Returned To The Lab! [Re: hamloaf]
#21920698 - 07/09/15 04:13 PM (8 years, 6 months ago) |
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Fantastic information
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Re: Sterile Equilibrium Returned To The Lab! [Re: TravelAgency]
#21923014 - 07/10/15 03:13 AM (8 years, 6 months ago) |
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Just happy to be back at it man. All the jars in the OP turned into bags 7-9-2015. 18 quart sized grainmasters were inoculated to 8 quarts of freshly sterilized rye in spawn bags. 1 jar to 1 bag. That's a 1:7 spawn ratio.

These bags were inoculated with grainmasters that have been fully colonized for well over a month. These jars also lived through the great contam storm of 2015. Not much hope is held, but as many provisions to sterility that could be thought of were employed before even opening the cultures. Attempting to expand grainmasters seemed like a much better option than throwing this particular round of spawn away. All the jars smelled good, so it'll be seen what culminates by the middle of next week.
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blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Sterile Equilibrium Returned To The Lab! [Re: bodhisatta]
#21923152 - 07/10/15 05:22 AM (8 years, 6 months ago) |
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Quote:
bodhisatta said:
Quote:
hamloaf said: Yea, man. No problemo. A couple of things that were forgotten to be mentioned. As measures to maintenance of the lab's sterile equilibrium the flowhood is set to a timer to come on for two hours a day. Once at noon for an hour, then once at midnight for an hour. Also, all surfaces of lab including shelf tops are wiped once a week thoroughly with an alcohol soaked paper towel.
have you seen the micro droplet disinfectant technology yet? you can peroxide fog a room to dramatically reduce contam loads and get near sterile environments but you don't want to accidentally lolercaust yourself
as long as the pH of your bleach solution is at or near 7 it will be quite effective for immediate use.
what % is the peroxide they use to fog?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Re: Sterile Equilibrium Returned To The Lab! [Re: blindingleaf]
#21988726 - 07/24/15 07:20 AM (8 years, 6 months ago) |
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Sparing you the gory details, just wanted to drop by and report that this endevour did not work out. Temp ranges in the lab got out of control and were sustained there for 2 weeks thus ALL 18 bags broke out with bacteria and mold. Temps were at 90-100F.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Sterile Equilibrium Returned To The Lab! [Re: hamloaf]
#21988944 - 07/24/15 08:48 AM (8 years, 6 months ago) |
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How did mold get in the bags?
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
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Re: Sterile Equilibrium Returned To The Lab! [Re: bodhisatta]
#21988983 - 07/24/15 09:00 AM (8 years, 6 months ago) |
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The ambient temps getting too hot and being sustained slowed the mushroom mycelium down and encouraged the dormant invader spores to germinate.
Inoculated the grain masters like 3 months ago. Wasn't too shortly after the grain masters were inoculated that I was no longer granted access back into the lab for what turned out to be 3 months. During that time the lab's temp range got out of control and stayed in the 90's for the first few months. As the environment's ambient temperature raised so did the temps in the lab to the 95-100F range. Not to mention the contam storm that these jars lived through opened the vector for invaders to have caught a ride with mushroom mycelium and get transferred into the receiving bags.
There were 18 bags all together. Lost the first 9 before 30 percent colonization and the shake. The remaining 9 bags fell to contamination within 3-5 days of being shook.
Edited by hamloaf (07/24/15 10:13 AM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Sterile Equilibrium Returned To The Lab! [Re: hamloaf]
#21989008 - 07/24/15 09:08 AM (8 years, 6 months ago) |
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I think I missed the part about the contam storm. Are you thinking that the sporeload on your filters was simply too high and compromised all the masters?
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
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Re: Sterile Equilibrium Returned To The Lab! [Re: Pastywhyte]
#21989024 - 07/24/15 09:12 AM (8 years, 6 months ago) |
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yea, that or bacteria could have invaded through the threads of the jar and the media closure as well. Three months and a contam storm later opens several vectors for contamination to invade. Couple that with the extremely high temps and forget about it.
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