|
 Exiledmonkies
Peaceful Warrior
Registered: 12/04/11
Posts: 511
|
 Steps to avoid bacterial contamination? 
#16354344 - 06/09/12 01:07 AM (11 months, 4 days ago) |
|
|
I've been doing rye jars for spawn for months but 3/10 jars I keep getting hit with bacteria leading to yellowish sweet smelling liquid that gathers on the bottom of jars making them useless. I've had great results with the other jars.
Steps that I'm using.
-Soaking and rinsing the rye before boiling.
-Making sure the jars are in the pressure cooker for at least 90min at 15PSI.
-If I reuse jars making sure they are cleaned with bleach and boiling water before they are put in the PC
-Working in front of a flowvent when doing G2G transfers. I thought maybe the filters on the flowvent might be contaminated so I swapped them out for new but that wasn't the problem.
I make sure to use elbow high medical gloves + rub alc + a organic vapor mask.
It can't be a contaminated syringe because I'm using healthy colonized jars for G2G as shown below. I also maintain a very clean house and workspace.
Healthy jars-
Jars contaminated with bacteria (the same and only problem I've had during colonization)
Ty for all input!
|
VaeVictum
Sativa Cyborg
Registered: 06/01/11
Posts: 4,397
|
Re: Steps to avoid bacterial contamination? [Re: Exiledmonkies]
#16354349 - 06/09/12 01:09 AM (11 months, 4 days ago) |
|
|
Hope you get someone who helps you out with this and pin points where the issue is coming from. Bacterial contamination is my least favorite of them all. 
VV
|
Exiledmonkies
Peaceful Warrior
Registered: 12/04/11
Posts: 511
|
Re: Steps to avoid bacterial contamination? [Re: VaeVictum]
#16354377 - 06/09/12 01:15 AM (11 months, 4 days ago) |
|
|
Likewise...I thought it might be to high of a temp or rh but I adjusted those but that wasn't the problem either. I keep all my dry unused rye in a airtight bucket as well. Mold is easy to prevent but this problem is just one big headf**k...
|
K1ngSp4de
CHUT UP!!!
Registered: 01/16/12
Posts: 561
Loc: 
|
Re: Steps to avoid bacterial contamination? [Re: Exiledmonkies]
#16354413 - 06/09/12 01:23 AM (11 months, 4 days ago) |
|
|
If the organic vapor mask is the one I found on google, the painters mask made by 3M, that could be the problem. Those masks are made to protect you from the environment, not to protect your work from you. For this you use surgeons masks.
--------------------
If a law is unjust, a man is not only right to disobey it, he is obligated to do so.
- Thomas Jefferson
CHANGE=CHANGA vowels don't exist in hyperspace.
Si peccasse negamus fallimur et nulla est in nobis veritas.
|
Exiledmonkies
Peaceful Warrior
Registered: 12/04/11
Posts: 511
|
Re: Steps to avoid bacterial contamination? [Re: K1ngSp4de]
#16354575 - 06/09/12 02:03 AM (11 months, 4 days ago) |
|
|
Its not the on made by 3M its a real LAB mask I acquired through my food science/tech courses. It has the same effect as a surgeon's mask but has a double charcoal filter chambers which I replace once a month.
I also take a extra step of showering + brushing teeth + mouthwash before any contact/work with my experiments.
|
consumesomeshrooms
( . )( . )
Registered: 02/21/11
Posts: 2,855
|
Re: Steps to avoid bacterial contamination? [Re: Exiledmonkies]
#16354680 - 06/09/12 02:40 AM (11 months, 4 days ago) |
|
|
Quote:
Exiledmonkies said:
I also take an extra step of showering + brushing teeth + mouthwash before any contact/work with my experiments.
also even if you use the cleanest of procedures you'll most likely have about a 10% contam rate
Edited by consumesomeshrooms (06/09/12 02:42 AM)
|
 i GrOw StUFF
Stranger
Registered: 01/14/11
Posts: 1,694
Loc: Space
Last seen: 3 months, 25 days
|
Re: Steps to avoid bacterial contamination? [Re: Exiledmonkies]
#16354708 - 06/09/12 02:54 AM (11 months, 4 days ago) |
|
|
My take is this (just my opinion): Most teks out there of grain preparation are great except for one thing, the drying time after boiling/simmering. I've learned that following some of those teks to the T of letting grains drain in a strainer for a little and loading into the jars is bull and never works out for me. I bet that those jars before G2G were subject to bacterial contam BEFORE the transfer. I've learned that myc likes dryer jars as opposed to wet jars. What I do is after I simmer, I lay those grains out on a towel (out of the sun and no fan) until they are dry on the outside, hence THE TOILET PAPER TEST...So 12-24 hour soak, simmer, dry.....sometimes I dry mine up to 12 hours....I bet thats the problem barring your PCing skills aren't up to par. When ever I got bacterial contam froma clean culture its been because of my grains being too wet.
--------------------
The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.
|
i GrOw StUFF
Stranger
Registered: 01/14/11
Posts: 1,694
Loc: Space
Last seen: 3 months, 25 days
|
|
Quote:
consumesomeshrooms said:
Quote:
Exiledmonkies said:
I also take an extra step of showering + brushing teeth + mouthwash before any contact/work with my experiments.
also even if you use the cleanest of procedures you'll most likely have about a 10% contam rate
No way, if you're getting 10% contams AND you're experienced you must be doing something seriously wrong.
--------------------
The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.
|
Exiledmonkies
Peaceful Warrior
Registered: 12/04/11
Posts: 511
|
Re: Steps to avoid bacterial contamination? [Re: i GrOw StUFF]
#16354738 - 06/09/12 03:05 AM (11 months, 4 days ago) |
|
|
Quote:
i GrOw StUFF said:
My take is this (just my opinion): Most teks out there of grain preparation are great except for one thing, the drying time after boiling/simmering. I've learned that following some of those teks to the T of letting grains drain in a strainer for a little and loading into the jars is bull and never works out for me. I bet that those jars before G2G were subject to bacterial contam BEFORE the transfer. I've learned that myc likes dryer jars as opposed to wet jars. What I do is after I simmer, I lay those grains out on a towel (out of the sun and no fan) until they are dry on the outside, hence THE TOILET PAPER TEST...So 12-24 hour soak, simmer, dry.....sometimes I dry mine up to 12 hours....I bet thats the problem barring your PCing skills aren't up to par. When ever I got bacterial contam froma clean culture its been because of my grains being too wet.
I thought about this. My method is very close. A couple hours before I work with the grains I spray the entire house with Lysol neutra air. After simmering the grains (checking on them every minute so they don't get over hydrated) I place them in a long open tub layered with clean papertowels and let them air dry till cooled. So there is no moisture that lines the jars when the grain is placed in them. I then check each jar carefully for split grains or anything that shouldn't be there. I always put them in the PC for a min of 90min.
I found that removing the foil from the jars after they were in the PC is a bad idea because it causes excess moisture to gather inside the jar as it cools. I wait till they cool then use them ASAP. I never let a jar go unused for more then 2days.
|
consumesomeshrooms
( . )( . )
Registered: 02/21/11
Posts: 2,855
|
Re: Steps to avoid bacterial contamination? [Re: i GrOw StUFF]
#16354753 - 06/09/12 03:13 AM (11 months, 4 days ago) |
|
|
not necessarily pretty much every grow i've done there's always that one or two jars that decide to turn on me. a completely contaminate free grow for a mushroom cultivator is kind of like getting a strike for a drunk bowler at a birthday party.
|
i GrOw StUFF
Stranger
Registered: 01/14/11
Posts: 1,694
Loc: Space
Last seen: 3 months, 25 days
|
|
well, what kind of PC do you have? Are you letting the PC pump out plenty of steam before you start building the pressure? Are you doing the toilet paper test? Which is awesome....How long are you letting the grains dry for?....Its not about cooling them, its about drying them so that there is no shining glistening on their outsides.
--------------------
The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.
|
i GrOw StUFF
Stranger
Registered: 01/14/11
Posts: 1,694
Loc: Space
Last seen: 3 months, 25 days
|
Re: Steps to avoid bacterial contamination? [Re: i GrOw StUFF]
#16354796 - 06/09/12 03:31 AM (11 months, 4 days ago) |
|
|
oh and there is no need to be crazy clean while preparing the grains. Don't be spraying all types of things around them while preparing them. Spraying things around the clean room is for glovebox things when you're inoculating....
--------------------
The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.
|
Exiledmonkies
Peaceful Warrior
Registered: 12/04/11
Posts: 511
|
Re: Steps to avoid bacterial contamination? [Re: i GrOw StUFF]
#16354850 - 06/09/12 03:50 AM (11 months, 4 days ago) |
|
|
I just use the spray as a extra step I never spray to soon before working so the grains or w/e aren't making contact with the chemicals. There are also ion air filters placed around the house which are on whenever I'm not working on any mycology experiments.
I use a cheap PC but it does reach 15PSI. I let the grains dry enough so they don't have any gloss to them but still look hydrated. *So there is no moisture on the outside of them at all.
|
consumesomeshrooms
( . )( . )
Registered: 02/21/11
Posts: 2,855
|
|
Quote:
consumesomeshrooms said:
Quote:
Exiledmonkies said:
I also take an extra step of showering + brushing teeth + mouthwash before any contact/work with my experiments.
also even if you use the cleanest of procedures you'll most likely have about a 10% contam rate
i just realized that i gave you your own advice op my bad
|
deucedbi9
Stranger
Registered: 10/24/06
Posts: 1,151
Loc: UK
Last seen: 4 hours, 3 minutes
|
Re: Steps to avoid bacterial contamination? [Re: Exiledmonkies]
#16354971 - 06/09/12 04:50 AM (11 months, 4 days ago) |
|
|
This "flow vent" you are working in front of. Is it a properly constructed laminar flow hood, or something you bought from a shop? Have you tested it? For instance, uncover an agar plate in front of it and see if you get a mold or bacterial growth.
--------------------
Cheap coir in the UK from £0.88p (inc p+p) per brick. Link
whether low pressure sucks or high pressure blows...
it's a bugger to cycle in
|
Exiledmonkies
Peaceful Warrior
Registered: 12/04/11
Posts: 511
|
Re: Steps to avoid bacterial contamination? [Re: deucedbi9]
#16354996 - 06/09/12 05:05 AM (11 months, 4 days ago) |
|
|
Quote:
deucedbi9 said:
This "flow vent" you are working in front of. Is it a properly constructed laminar flow hood, or something you bought from a shop? Have you tested it? For instance, uncover an agar plate in front of it and see if you get a mold or bacterial growth.
Its a properly constructed Flow hood I built myself and I have tested it exactly the way you mentioned. It works just fine. I've been calling them Flowvents since the noob days and still havent broken that habit lol...
|
deucedbi9
Stranger
Registered: 10/24/06
Posts: 1,151
Loc: UK
Last seen: 4 hours, 3 minutes
|
Re: Steps to avoid bacterial contamination? [Re: Exiledmonkies]
#16355035 - 06/09/12 05:45 AM (11 months, 4 days ago) |
|
|
Sorry geezer. Had to ask.  It was the use of the term "flow vent" that led to the question.
The only other question that comes to mind is, what are you inoculating the grain masters with. Wedges, GLC, I'm guessing not spores, or, LC 
May be your batch of rye has a particularly virulent and hardy species of bacterial endospore.
I've never used rye, trying to source some atm. I use wheat grain, same family as rye, so I read. Though it's cleaner than WBS, it doesn't perform as well IME.
Never had a bacterial contamination with either though.
I think the UK.GOV, in it's imperious and omnipotent wisdom, banned endospores from our shores.
Either that, or it's the 'cooking' method I use.
Made up four jars yesterday. Started at 7.00am, pasteurised grain for an hour, strained/rinsed/dried and jarred by 8.50am finished the PC run at 10.20am.
Inoculated at~3.30pm. Passed experience tells me that these will complete without contamination.
Hope it goes better for you in the future.
Good luck.
--------------------
Cheap coir in the UK from £0.88p (inc p+p) per brick. Link
whether low pressure sucks or high pressure blows...
it's a bugger to cycle in
|
TheEaglesGift
The Monolith
Registered: 04/11/11
Posts: 3,288
Loc: Amsterdam, Netherlands
Last seen: 1 month, 24 days
|
Re: Steps to avoid bacterial contamination? [Re: deucedbi9]
#16355045 - 06/09/12 05:56 AM (11 months, 4 days ago) |
|
|
"a completely contaminate free grow for a mushroom cultivator is kind of like getting a strike for a drunk bowler at a birthday party."
I totally disagree. I've had many contamination free grows. I don't settle for mediocrity. If you are having trouble with contamination, you have two option: 1) Accept a certain margin of error, which is fine if you are okay with that or 2) isolate the issue causing contamination and rectify it.
Personally I believe in striving for 100 percent success rate. Obviously the occasional mold spore, chance and human error will intervene, but I still strive.
--------------------
Edited by TheEaglesGift (06/09/12 05:58 AM)
|
Exiledmonkies
Peaceful Warrior
Registered: 12/04/11
Posts: 511
|
Re: Steps to avoid bacterial contamination? [Re: deucedbi9]
#16355046 - 06/09/12 05:56 AM (11 months, 4 days ago) |
|
|
My masters are usually inoculated with a agar wedge from there its always G2G. Every once and awhile I use MS syringes but thats rare since I've been enlightened by the experts that using MS is a bad idea.
I've always used the same type of rye 100% organic. So maybe that is the problem. I would guess if it was the rye they would all fail though. I'll try a few other brands and maybe WBS.
The only other thing I can mention (which is annoying as hell) is the contamination doesn't take effect till the jar is 75%+ colonized. Then the yellow liquid really begins to form and stalls the myc growth.
A couple of the jars had the yellow liquid and 100% colonized so I popped them open smelled them (smelled fine) so then I used them in a minimono grow. Complete fail...after opening up the lid days later to check on it the smell was so sour I almost vomited. Removed that from the house ASAP and cleaned for the next few hours.
|
deucedbi9
Stranger
Registered: 10/24/06
Posts: 1,151
Loc: UK
Last seen: 4 hours, 3 minutes
|
Re: Steps to avoid bacterial contamination? [Re: Exiledmonkies]
#16355089 - 06/09/12 07:17 AM (11 months, 4 days ago) |
|
|
Quote:
Exiledmonkies said:
I've always used the same type of rye 100% organic. So maybe that is the problem. I would guess if it was the rye they would all fail though. I'll try a few other brands and maybe WBS.
Yes it's perplexing.
The wheat I use isn't even food grade (pet shop) let alone organic.
When I used WBS, using the cooking method I referred to, I'd dry it one strainers worth at a time...I can only PC four jars at a time so it's not that time consuming... in a roasting pan with my daughters hair drier   until it reached this stage>Link
You've probably seen and know all this 'Em', others may not.
With wheat, I rinse and strain out as much water as possible, with a sort of 'dropping' action, and dump onto doubled over sheets of newspaper in the roasting pan.
Change the newspaper a couple of times and don't leave it to long at first or it will soak and tear as you lift it to place a dry piece underneath. Takes about three changes of paper using the dry part of each previous one to push the grain around to help dry it.
Pour the now dry, as per above link, straight into the jar from the paper after sprinkling it with gypsum. Pro-tip: Keep an old 'spice jar' which has holes in the lid full of gypsum for this purpose.
Good luck.
I'll just add that, before pc'ing I always wipe the rim of the jar down with a wet cloth, then dry, like the PF tek'ers do their jars. It's one of my more anal 'things'. 
--------------------
Cheap coir in the UK from £0.88p (inc p+p) per brick. Link
whether low pressure sucks or high pressure blows...
it's a bugger to cycle in
|
|
Images only 
Threaded
Previous
Index
Next
Shop for: 
Edit 
Reply 
