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Amazon Shop for: ½ Pint Jars, HEPA Filter, Pressure Cooker, Rye Grain, Wild Bird Seed

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OfflinePugslee_Atoms420
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Registered: 08/20/02
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never had problems before, but having them now!!!
    #1139370 - 12/13/02 06:46 PM (14 years, 3 months ago)

Hey, I was just wondering if anyone could give me some idea as to why my jars are contaminating now and they haven't before. I have been getting a lot of this yellowish dusty looking particles that seem to contaminate entire hars within days. The only thing I am doing differently is using polyfill lids as per the Anno tek, whereas before I was usually using MicronMagik filters and injecting right through the filter. I always use the oven tek and lysol a bunch beforehand; but now because of the new lids I have been just opening them a bit quickly and innoculating through the opening. I am getting problems with almost every rye jar, and every single bird seed jars no matter how nice they look when first out of the p/c. So far as I know everything is cool with my incubators too, they always worked before. I am using a new Presto pressure cooker, but I have checked it and it is working properly. Well here are the pics, let me know if anyone has any ideas!!!! Thanks
[image][/image]
[image][/image]

p.s. These 2 jars were spawned with a heaping tablespoon of Maz rye spawn


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OfflineAnnoA
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Re: never had problems before, but having them now!!! [Re: Pugslee_Atoms420]
    #1139662 - 12/13/02 10:23 PM (14 years, 3 months ago)

> I have been just opening them a bit quickly and innoculating through the opening

Obviously the air in your kitchen is very mold-spore laden.


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InvisibleTinMan
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Re: never had problems before, but having them now!!! [Re: Anno]
    #1140197 - 12/14/02 06:50 AM (14 years, 3 months ago)

Give your house a good sterilization and spray the air down with alcohol.


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OfflineSuntzu
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Re: never had problems before, but having them now!!! [Re: Pugslee_Atoms420]
    #1140202 - 12/14/02 06:53 AM (14 years, 3 months ago)

I'd predict that your success rate would improve again if you went back to the polyfill;  Also, you'd probably enjoy the same success if you did the polyfill inoculations on your kitchen counter vs. inside the oven.  You'd likely have a higher success rate with the filter disks if you moved to a simple glovebox system.  As nasty as it looks, this unit has served me flawlessly for years:



I know many people say the oven tek works for them;  In order to convince me it's making any difference, they'd need to show me that doing inoc's on the kitchen counter do NOT work, and moving into the oven DOES work.  The concept is fundamentally flawed, IMO, and proported 'successes' are masked by lack of comparison.    People who regularly succeed with oven tek inoc's should try doing it on the kitchen counter.  I'd wager they'd see similar results [but then again, I've lost many a wager :wink: ].

Polyfill may be your best bet;  Any time you unscrew and remove a grain jar lid, you should be in *that* state of mind where you are personally clean, your hand movements are pre-planned out, and operations are performed quickly.  When done correctly, such inoculations can occur in less than a second.  There is a lot more of a fidge factor when you don't remove the lids, as per your polyfill experience.

 


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OfflinePugslee_Atoms420
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Re: never had problems before, but having them now!!! [Re: TinMan]
    #1141820 - 12/14/02 10:31 PM (14 years, 3 months ago)

Okay, thanks everyone for the comments. I really do think that it may be the kitchen; any suggestions how I should thoroughly clean everything? 10% bleach spray or higher? Anything besides that, like alcohol spray mentioned by? If an alcohol spray would be better to sanitize or whatever, what concentration of alcohol to water should I use? I will defintely also try doing things on the counter top for the next batch, and then I figure it would be wise to finally break down and build a glovebox. Sunzu, was wondering about your glovebox and how you made it---- like what kind of gloves and such you use to seal or whatever. Also what kind of spray you use to sterilize it. I need to definitely get cracking on building that..... Would you mind if I pm'd you Sunzu to get some pointers on how you made/use yours? Or does anyone have a good proven tek for a glovebox that holds around 6-10 Quarts? Thanks everyone!!! Peace


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OfflineMadcapMagician
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Re: never had problems before, but having them now!!! [Re: Pugslee_Atoms420]
    #1151393 - 12/18/02 10:54 AM (14 years, 3 months ago)

You need 70% alcohol. THIS IS IMPORTANT! If you use too high a percentage of alcohol you simply cause any bacteria/mould present to form spores (I think because of the dehydrating action of the alcohol) which will then grow like crazy in your nicely prepared medium! Also, when you use 70% IMS (industrial methylated spirits - I use it in a cytotoxics manufacturing unit for surface sterilisation) make sure it doesn't pool, make sure you get all surfaces wet, but most importantly, allow everything to dry - 70% IMS kills bugs as it evaporates, so using stuff that's soaking wet, albeit with alcohol, is about as useful as a chocolate fireguard! You need to clean down your working area first by spraying it with IMS. Once the worksurface is dry, spray down all your equipment and put it on the clean work surface. Once it's all dry, use it immediately. If you wear gloves while you work, spray your hands before you put the gloves on, then spray the out side of the gloves as and when necessary.


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OfflineSeussA
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Re: never had problems before, but having them now!!! [Re: MadcapMagician]
    #1154637 - 12/19/02 10:45 AM (14 years, 3 months ago)

cause any bacteria/mould present to form spores

This is incorrect! Bacteria do not form spores. Some bacteria, such as the various Bacillus species, produce endospores, as a survial response, but not as a reproductive mechanism. Endospores also take a while to form... longer than it takes for the alcohol to do its work.


--------------------
Just another spore in the wind.


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OfflineSuntzu
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Re: never had problems before, but having them now!!! [Re: Pugslee_Atoms420]
    #1157853 - 12/20/02 12:33 PM (14 years, 3 months ago)

I PM'd you with some of the basics, but I thought it would be good to mention in this thread. . .

It is not nearly as important to have things 'sterilized' by lysol, alcohol, etc., as it is to have the air clean when doing these inoculations.  Your inoculum was made before, or by someone else; the jars are fresh from the PC;  Depending on what you are inoculating with and how, you could get away with just swabbing the inside of the glovebox out with WATER to dislodge any mobile dust particles [honestly, I use those Chlorox antimicrobial wipes, but I'd bet even they aren't necessary].  Any utensil that might be used in the transfer [scalpel, exposed needle, spoon, watever] should be swabbed with lysol/alcohol/bleach/lighter flame, of course;  But don't needlessly expose your lungs to large quantities of this stuff. 

Instead, get a small but decent hepa filter [walmart-types are fine]. It's best to use a small room that you can close the door, sealing it off somewhat.  I use my bedroom, and do not go crazy cleaning it up beforehand.  Turn off the house heater so no new air is being pumped in.  Have dirty clothes, any major dander sources picked up.  Turn on the hepa filter ~15-30 minutes before you will be ready to inoculate.  Have the glovebox swabbed, jars/inoculum in position within.  Close the door to the room, go smoke a doob for ~15 minutes, take a shower;  Fresh from the shower, enter the pseudo-clean room and go straight to the glovebox.
This works incredibly well for a lysol/alcohol-less procedure.  No latex gloves, nothing crappy for the lungs.  *edit*  [except the doob]

I used to be a lysol madman, BTW, and it does work;  Just read the ingredients in that can before saturating your breathing space with it :wink: 


Edited by Suntzu (12/20/02 12:35 PM)


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Amazon Shop for: ½ Pint Jars, HEPA Filter, Pressure Cooker, Rye Grain, Wild Bird Seed

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