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OfflineChemical_Smile
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Whats actualy going on in my house?
    #1548450 - 05/14/03 04:44 PM (13 years, 6 months ago)

I want to know because despite all my carefull attempts to avoid contams I still get them like 50% of the time.  I need to know how the various nasty things get into my jars and casings.  Lets asume that all fans, vents, etc are off and I have misted everything w/ alcohol.  If I could see microscopic contams in my house would they be swirling around the air like a sand storm or slowly flying by like a light snow?  Would they be on objects (tools,me, wall, etc) and drop off into my jars or do the contaminated surfaces actualy have to contact my jar (like getting sick from touching a public phone)?  I always mist and always use a glovebox so I am stumped as to how I am getting contaminated.  I have arms built into the glovebox but there is a small gap where I stick my hands (gloved and sprayed w/ alcohol) through.  Unless the nasty stuff was sitting there just waiting to get my jars like the pet cat that goes flying out the door the second you open it I dont see how I have problems.  ( I presoak my grain 12 hours before I boil it and pc)    :confused: 


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OfflineMAIA
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Re: Whats actualy going on in my house? [Re: Chemical_Smile]
    #1548776 - 05/14/03 06:31 PM (13 years, 6 months ago)

Your missing as possible cause for jars: how long inside the PC, PC pressure, syringe sterility, flaming the needle between jar inoculations, lids tight or loose, holes untapped (if any) ?

As for casings goes, do you keep your terrarium in a clean place ?
You are a "taxi" for contams, change cloth to something clean before working. Also this time of year has a very high spore counting, so you can also consider that.

When using the glove box, which kind of gloves do you use ?
Besides, you should always uncover your arms till your elbows and wash both arms very well before working in the glove box.

MAIA


--------------------
Spiritual being, living a human experience ... The Shroomery Mandala



Use, do not abuse; neither abstinence nor excess ever renders man happy.
Voltaire


Edited by MAIA (05/14/03 06:33 PM)


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OfflineChemical_Smile
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Re: Whats actualy going on in my house? [Re: MAIA]
    #1548823 - 05/14/03 06:53 PM (13 years, 6 months ago)

Presoaked rye for 12 hours to let contams germinate
Boiled Rye till it was swollen but not bursting
Put in the pc for 1 hour 10 minuites at 15 psi
Wiped down everything inside and out w/ alcohol
washed hands put on latex gloves then more alcohol
(In glovebox) heated syringe needle until all parts had glowed red
cooled w/alcohol on paper towel.
Undid lids and squirted in spores then put filter disk back on w/band to hold it
Sprayed air again
Removed from glove box
Put foil over tops of jars
placed jars back in original box (after spraying it too)
Placed in closet.
Lost over half to green (one had pink too)  :confused: 


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Anonymous

Re: Whats actualy going on in my house? [Re: Chemical_Smile]
    #1548906 - 05/14/03 07:36 PM (13 years, 6 months ago)

I think its possible that some areas of the world just have more spores in the air than others....I barely get any contams even when not taking all the recommended precautions


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OfflineChemical_Smile
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Re: Whats actualy going on in my house? [Re: ]
    #1549067 - 05/14/03 08:56 PM (13 years, 6 months ago)

Yeah, somethin about my attempts attracts em. It might be high spore count in my grain, I thought of that though and thats why I soak it beforehand. Anybody know how green mold gets spread? Its airborne right?


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OfflineSeussA
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Re: Whats actualy going on in my house? [Re: Chemical_Smile]
    #1550297 - 05/15/03 07:54 AM (13 years, 6 months ago)

Quote:

Boiled Rye till it was swollen but not bursting
Put in the pc for 1 hour 10 minuites at 15 psi




What I am about to describe is normally not needed, but you have problems so lets try to find the source.  Let the jars cool and PC them again 24hrs later for the same time at the same pressure.  If you are looking for the source of your problems, only PC half the jars a second time.  Be sure you mark which jars got the double kiss in the PC.  If one set of jars contaminates, and the second doesn't, then you know this is the source of your problem.

Be sure your have everything in the PC that will be part of your jars.  The substrate, the lids, rings, filter disks, foil that you use to cover them, etc.

Quote:

Wiped down everything inside and out w/ alcohol




Be sure the alcohol is about 30% water, or 70% alcohol.  It also helps to spray a generous amount of lysol into the glove box and around the room before you work.  Lysol, alcohol, and flames don't mix, so be careful.

Quote:

washed hands put on latex gloves then more alcohol




Take a good long hot shower before this step.  Use lots of soap and scrub down like a madman.  Use a clean towel and put on clean clothes.  Wear a short sleeve shirt.

Quote:

(In glovebox) heated syringe needle until all parts had glowed red
cooled w/alcohol on paper towel.




I would skip the cooling with alcohol on a paper towel.  After heating the needle, just chill for a bit while it cools down from red hot.  A small squirt of your spore solution through the needle will help.  It will hiss and kill a lot of your spores, but you wont pick up anything either.  Repeat the above step between each jar that you innoculate, but not each hole within a single jar.

Quote:

Undid lids and squirted in spores then put filter disk back on w/band to hold it




Time is money, keep those lids off as little as possible.  Are you using a vermiculite seal on top of the substrate?  If not, add one... you will have to poke the needle through the verm layer to get the spores into the substrate.  Oh wait, you are using rye berries, never mind on the verm seal.

Quote:

Sprayed air again




Not needed at this point.  Doesn't hurt to wipe the jars down with alcohol though.

Quote:

Put foil over tops of jars




You are using filter disks, correct?  Be sure the foil doesn't prevent the jars from breathing.  Some contams thrive when the air inside the jars becomes stagnent.

Quote:

Placed in closet.




Did you spray down the closet as well?


Here are a few additional thoughts...

90% of all contamination lives within one foot of the floor (gravity pulls them down).  Work on a table, not on the floor.  Avoid moving around too much or having fans run or anything else that can stir up the floor contam.  Store your jars on a shelf in your closet, not on the floor.

Before you start working do everything you can to reduce the spore load in your house.  Open the windows up for a few hours and air the sucker out.  Wipe down the hard surfaces with cleaners.  Vacuum any carpets, curtains, couches, etc.  Change out the filters in your furnace.  Run an air filter in your work area.  Spray lots of lysol into the air.

About two hours before you start working, lock everything down tight.  Close all the windows, turn off the AC or furnace, turn off any fans, etc.  You want to give time for anything suspended in the air to settle down.

Use a different spore syringe.  The one you are using may be contaminated.

Don't open the jars or handle them once you put them up in storage.  Check them once a week or so and toss any that show contamination.

Use fresh jars... do not reuse contaminated jars, rings, or filter disks until you know that this is not a source of contamination.

Try using less substrate and smaller jars.  The smaller the jar, the faster it will colonize and the less time contamination will have to move in.

Never open a contaminated jar anywhere near where you will be working in the future.  Best to just toss them in the trash without opening at all.

Try switching back to pf style cakes if rye berries are giving you trouble.

With rye berries, you want to remove the jars from the PC as soon as you can, while they are still very hot, and shake them up.  If you don't, you will end up with soggy berries at the bottom, dry berries at the top, and usually contamination.

The contamination is coming from somewhere.  Look at everything you are doing and try to figure out if it is a risk or not.  Change your procedures slowly over time until you know where the problem was.  Once you get it down, follow the same procedure again and again.  Never assume something is clean.  :smile:

Hope this helps.  Good luck!


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


Edited by Seuss (05/15/03 07:56 AM)


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OfflineChemical_Smile
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Re: Whats actualy going on in my house? [Re: Seuss]
    #1550798 - 05/15/03 01:26 PM (13 years, 6 months ago)

Good stuff, thanks.  :grin:  I have a better idea of how the enemy spores are behaving now.  Im gonna try double pcing like you said just to make sure.  It might be like digging him up to cut off his head but at least I will be able to rule out that possibility.  Do I have to worry about the 2X jars drying out?  I don't quite get it though, I thought that properly pressure cooking it for the right amount of time killed EVERYTHING.  Is that not 100% true?  I had considered just doing filter bags full of compost and skipping rye all together.  Just 1 or 2 holes from the needle while under a alcohol pad and then imediate super glue patching of the holes.  Thing is thats alot of substrate to have to p.c.  If its not 100% effective it could be a problem. 

How trustworthy are filters?  Previously I had just used the mm filter disks on top but went to the foil to be xtra safe.  Could contams possibly get through?  I always used lysol or alcohol on the outside of the filters when I had to open them for grain to grain transfers.  I had wanted to cold shock some casings in the fride seeled up in bags but that is mean fucking neighborhood as far as contams so these filters people are selling would have to be 100%.  Anyways Im rambling, thanks for the help!  :smile: 


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OfflineMAIA
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Re: Whats actualy going on in my house? [Re: Chemical_Smile]
    #1551726 - 05/15/03 06:06 PM (13 years, 6 months ago)

After reading Seuss excellent post i have two thoughts about this.
Broken PC. Does it have a gauge or something ? Is it reading well ?
Filter discs. Don't use filter discs in one or two jars next time you try, so you can control that aspect too.

MAIA


--------------------
Spiritual being, living a human experience ... The Shroomery Mandala



Use, do not abuse; neither abstinence nor excess ever renders man happy.
Voltaire


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OfflineSeussA
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Re: Whats actualy going on in my house? [Re: Chemical_Smile]
    #1551865 - 05/15/03 06:34 PM (13 years, 6 months ago)

As long as your PC doesn't dry out, your water content should be fine. That is why we use a PC instead of dry heat sterilization.

As far as timing goes on the PC, think of ice melting as an analogy. If you have a little ice cube (or a little jar in your PC) it doesn't take very long for the ice to melt. If you have a large block of ice (or a large jar in your PC) it takes a lot longer for the ice to melt. It all depends upon the amount of mass that you are trying to bring to a certain temperature. The more you put in there, the longer it takes to ensure that everything is evenly heated.

Rye berries are also a lot larger than bits of flour. It takes a lot longer for the heat to penetrate to the center of the berry. I use a professional autoclave and cook my berries one single time for 1.5 hrs at 25 psi (aprox 265 F) and don't have any problems. I simmer the berries as you describe before hand. If you are at a high altitude, you will need to increase your cook times to make up for the difference in pressure. I am pretty much at sea level, but I used to live in the mountains and would double cook as I described above to compensate.

I have found that using filter disks without most of their area covered caused the top berries in the jar to dry out. In your shoes I would continue to cover with foil, but poke some air holes in the foil for ventilation. The foil isn't there to stop contamination as much as it is to stop excessive evaporation during colonization.

Unless your PC is faulty, I really doubt that the double cook will help much. If it does, to speed things up in the future, try a single cook at 1.5 hrs with berries. I had a hell of a time with bacillus and rye berries from poor sterilization times of the berries, but have never had a problem with green during colonization, so I am shooting in the dark a bit here.

When your friend tries again, let me know how it goes. Pay attention to detail and your friend should have no problems.

Edit: I forgot to address the PC killing everything... they have found bacteria living inside the liquid sodium inside the primary cooling loop in nuclear reactors. A single bacteria can reproduce once every twenty minutes. After just eight hours a single bacteria can multiply into a colony of millions.... by 24 hours this can increase to trillions of organisms. Your problem is more fungus related, but the idea still holds true. Just one single spore that lives through the PC cycle can ruin your crop... and there can be millions upon millions of enemy spores in there that need to be killed. You might kill 90% of all the spores in the first 10 minutes, and an addition 90% of what is left in the next 10 minutes, and so on... even after an hour at this rate you can still have a few spores left. It is easy to kill most of them, but difficult to kill all of them, and when it only takes one....

Also, remember that the more you handle the jars after sterilization, the more likely you are to transfer contamination to the outside of the jars. Some organisms are motile and will move around in search of food. The filter disk can stop the contamination, but if there is not a good seal between the disk and the glass, the contamination can still get through. Make sure those lids are locked down tiight. A wrap or two of parafilm around the rings and glass can be used to seal things up a bit tighter. Do a search on parafilm for the store bought equiv... I think anno posted once what you can use instead... some kind of plastic wrap if I remember correctly.

Always wash up, wear gloves, and use alcohol when handling your jars, even after they have been innoculated.


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


Edited by Seuss (05/15/03 06:43 PM)


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OfflineChemical_Smile
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Re: Whats actualy going on in my house? [Re: Seuss]
    #1552535 - 05/15/03 10:23 PM (13 years, 6 months ago)

Wow, thanks guys! I have honestly put alot of time and effort into this hobbie (still waiting on the payoff) and I realy appreciate your input.

Quote:

Broken PC. Does it have a gauge or something ? Is it reading well ?





Well Its a old one and my grandpa found in his attic, seems to be in good shape. Its kinda big, you can fit 7 quart jars in it w/ out stacking and almost (not quite) get another 7 on top of those. Has a guage on it and as far as I know it works. I dont know any symptoms of a faulty pc though except mayby it blowing up.

Quote:

I use a professional autoclave and cook my berries one single time for 1.5 hrs at 25 psi (aprox 265 F) and don't have any problems.




Thats way more cooking than I have done, not cause I mind waiting but thats what the tek said. Ive done 1 hour at 15. Now that I know it wont hurt anything Im deffinately gonna do them longer.

Quote:

Also, remember that the more you handle the jars after sterilization, the more likely you are to transfer contamination to the outside of the jars. Some organisms are motile and will move around in search of food. The filter disk can stop the contamination, but if there is not a good seal between the disk and the glass, the contamination can still get through. Make sure those lids are locked down tiight.




The more I think about it I am realy getting tired of seals and lids and all that. Like I said above I am thinking about just doing up big ass bags of shit/compost and skipping the grain part and just inoculate multispore through the bag. Even the best of the contams probably couldnt get through a few needle size holes before I can get some super glue and seal them up. How long would you recomend for the shit bags. Im using Large myco bags (18" high 8" wide 5" deep) and I want to fill them as full as possible while still alowing for room to mix the contents up. My thinking with this is that if I go big w/ my projects and even 1 of them works right the pay off will be worth it all. Composted manure and bags are both pretty cheep so I can afford to do up a bunch and even if most of them get contaminated I will still have plenty to work with. The time I would save not messing with grain would let me make alot more bags and syringes as insurance. Do you notice any flaws in my thinking on this?


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InvisibleCLuB99
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Re: Whats actualy going on in my house? [Re: Chemical_Smile]
    #1553833 - 05/16/03 01:23 PM (13 years, 6 months ago)

i'd test a sterilized jar leaving it in your incubator but without inoculating it, this way it's easier to track down the source of contamination


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InvisibleSixTango
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Re: Whats actualy going on in my house? [Re: Chemical_Smile]
    #1555735 - 05/17/03 06:48 AM (13 years, 6 months ago)

Sometimes simple things you overlook, cause contam's.

Once my foaf cleaned out some contam'ed jars. Doing so outside. A contam spore cloud developed in the immediate area.

Knowing those spores got all over him, before he entered his lil grow room, he showered & changed clothing.

Several days later, about 50% of his trays developed contams.

The FOOL finaly figured out that he wore the same  :crazy:baseball hat :crazy: - into his grow room that he was wearing in the aforesaid contam spore cloud.

That did it -- as sure as water is wet.

6T :wink:


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~whiskey river rafting, hot tubbing, dirty dancing & spending money on - wild women - having fun & just gonna waste the rest~


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