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OfflineSaryn
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Fighting contams, could use some advice
    #7726475 - 12/06/07 09:45 PM (16 years, 1 month ago)

Ok so I need some help figuring out exactly where in the process I am getting some tich contamination. For reference, the spots show up somewhat close to the dry verm layer. The contamination also only shows up about a week after inoculation.

First off, I've been trying to follow RR's video guide on youtube as precisely as I can. Rinse out the jars then proceed to fill with the BRF/verm mix. Before capping, I clean around the rims of the jar with a paper towel then add about half an inch of dry verm. Covered with aluminum foil, I then pressure cook the half pint jars @15 psi for 60 minutes once the pressure has been reached, then allow to cool for 30-45min before removing (mainly to allow the pressure to drop).

After at least 24 hours, I inoculate. Following the video, I use a rubber made style bin with holes cut for gloves. The whole bin is cleaned with alcohol and air sanitizer before use. I use latex gloves which are cleaned with alcohol before use. Following the inoculation procedure, I wipe the needle with alcohol, flame, and wipe again in between jars and also clean the jar lids with alcohol. Also, as per the guide, I leave the holes alone as it has been suggested that the dry verm layer works very well as a filter.

Where am I going wrong? I keep reading the guides and I'm not sure what I'm doing incorrectly or failing to do. Any help would be greatly appreciated!


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InvisiblePremedman1
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Re: Fighting contams, could use some advice [Re: Saryn]
    #7726529 - 12/06/07 09:58 PM (16 years, 1 month ago)

You follow very good sterile procedure. The next plausible explanation would be the syringe you're using to inoculate the jars.

What is the source of the syringe?


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OfflineSaryn
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Re: Fighting contams, could use some advice [Re: Premedman1]
    #7726553 - 12/06/07 10:01 PM (16 years, 1 month ago)

Got the syringes from spores101.com. I'm not sure if it was the syringes because I am doing many jars and its occurring over multiple strains.


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InvisiblePremedman1
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Re: Fighting contams, could use some advice [Re: Saryn]
    #7726625 - 12/06/07 10:19 PM (16 years, 1 month ago)

With that kind of sterile procedure, all I can surmise is the contamination is happening between the time the jars are cooked, and inoculation.

Suggestions are to let the jars cool overnight in the pressure cooker. When you pull out hot jars, the substrate and surrounding gasses in the jar shrink, pulling a substantial amount of air into the jars. This could be pulling contaminates in as well. The dry verm layer will not block everything. This could explain why the contaminates are appearing just below the vermiculite barrier.

Lastly, I would rubber-band a coffee filter over the lid. This would allow adequate gas exchange while preventing dust from getting to your layer of vermiculite, especially if it got moist during the PCing.


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Build a man a fire, he is warm for the night. Set a man on fire, he is warm for the rest of his life.


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OfflineSaryn
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Re: Fighting contams, could use some advice [Re: Premedman1]
    #7726635 - 12/06/07 10:22 PM (16 years, 1 month ago)

I was using micro pore tape to cover the holes prior to inoculation, are you suggesting to put on the filter after inoculation? Just wanted to clarify timing.


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OfflineNibin
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Re: Fighting contams, could use some advice [Re: Saryn]
    #7726687 - 12/06/07 10:37 PM (16 years, 1 month ago)

Shit man, you are really doing things properly.

RogerRabbit is saying that if micropore is used, and after injecting another strip is stuck over the needle hole, not even the dry verm barrier is needed.

Do you think your verm might have contam spores in it? or did you use a new bag?


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OfflineSaryn
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Re: Fighting contams, could use some advice [Re: Nibin]
    #7726696 - 12/06/07 10:41 PM (16 years, 1 month ago)

New bag, new everything. I am really running out of ideas on what is causing this. About the only possible thing I can think of is that when I check the jars, I somehow jar that top verm layer or something and allow what ever got in via air to shake down. Will be starting up yet another batch over the weekend, will try the coffee filters as well as any other suggestions.


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Invisibled4a2n0k
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Re: Fighting contams, could use some advice [Re: Saryn]
    #7726755 - 12/06/07 11:01 PM (16 years, 1 month ago)

I agree, let the jars cool in the PC.

You said you put on micro pore tape before inoculation? Do you then remove the tape or inject through it? I would think micro pore tape would be somewhat sterile out of the package, but other than that you can never be sure.

Just trying to cover all bases here.


--------------------
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Merrily, merrily, merrily, merrily,
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Offlineoxohawkoxo
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Re: Fighting contams, could use some advice [Re: d4a2n0k]
    #7726766 - 12/06/07 11:07 PM (16 years, 1 month ago)

Where are you putting on the tip of your syringe before inoculation? I had similar mystery contamination problem. Ever since I've changed to putting the tip on my syringes inside the still air box and after the air had time to settle, they've gone away. Might also try wiping the bottom of the needle portion and plunger/container portion with alcohol just as you put them on.

Putting your tape after inoculation is a good idea too. Putting it on before mean it's punctured when you let the jar sit and could be a vector though which contams are getting in. Hope someone's suggestions and idea here help. I understand how frustrating it can be.


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Invisiblethedefone
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Re: Fighting contams, could use some advice [Re: d4a2n0k]
    #7726782 - 12/06/07 11:12 PM (16 years, 1 month ago)

Try scrapping the verm barrier, maybe? Try it with tyvek between the lid and the screw ring before sterilization, and mark your inoculation holes so you're not stabbing all over the place. I also agree that leaving the jars in the PC overnight might help your success. Cooling them too rapidly may accidentally suck trich spores into your jars...

Who knows...


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OfflineCloneufc
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Re: Fighting contams, could use some advice [Re: thedefone]
    #7726864 - 12/06/07 11:42 PM (16 years, 1 month ago)

I too just got Trich in my BRF cakes. I made mostly manure cakes and a few BRF cakes. All my BRF cakes are contaminated. My manure cakes appear to be fine but its harder to tell because its alot darker.I must have made a dirty syringe. I think BRF get contams alot easier. Maybe you should try something else.


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OfflineSaryn
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Re: Fighting contams, could use some advice [Re: Cloneufc]
    #7727028 - 12/07/07 12:27 AM (16 years, 1 month ago)

Thanks for the comments!

As for the tape, I put it on before sterilization, then cover the jars with foil to keep the water out. I then inject through the tape without recovering for fear that contamination could get in off the tape. Ill try letting the cans sit overnight in the PC to see if that helps as well as tyvek between ring and lid.


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Offlineveda_sticks
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Re: Fighting contams, could use some advice [Re: Saryn]
    #7727438 - 12/07/07 05:32 AM (16 years, 1 month ago)

Definatly leave the jars in the pressure cooker overnight, as long as the lid is on the pressure cooker, its pretty sterile in there and there is very little chance of anything getting in.

Have your pressure cooker next to your glove box so you can transfer to glovebox quickly.

It sounds like your sterilisation procedures are pretty solid.

What was the moisture content of your substrate, was it too wet?? if it was its possible the verm filter got damp allowing contams to grow threw it.

I think your using the same tek as me, drinking glasses with tinfoil as lids, i have had great success with that. I dont bother with the micropore tape, but if you have it use it, you can leave it out when pressure cooker, when you inoc just peirce straight through the tinfoil then place a peice of tape over the hole.


--------------------

PF TEK - writeup by EvilMushroom666
Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD
Cakes can and will pin! - So you think cakes suck for pins. Your wrong
Franks Simple Coir/Verm Tek
Franks Proper Pasturisation Tek
Franks Spawning To Bulk - Monotub
Professor Pinheads RTV Injection Port Tek
Foo Mans No Soak WBS Prep Tek


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OfflineNibin
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Re: Fighting contams, could use some advice [Re: veda_sticks]
    #7727445 - 12/07/07 05:37 AM (16 years, 1 month ago)

He says in the first post he uses jars, plus lids, plus dry verm barrier, plus micropore.


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Offlineveda_sticks
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Re: Fighting contams, could use some advice [Re: Nibin]
    #7727454 - 12/07/07 05:42 AM (16 years, 1 month ago)

Ah, never mentioned puttin gthe lids on with the holes in it.

You can try the tyvek method, it works too and u dont need the verm filter, so you can get that little bit extra substrate in the jar.

Put the tyvek ontop of the metal lid with rubber seal, and put the ring on and tighen. Simply mark where your inoc holes are, wipe with alcohol before inoc and peirce through the tyvek, covering with micropore tape afterwards.

Its worth a shot, plus i believe you can get them for free at the post office if your in the US.

Other than that,im stuck


--------------------

PF TEK - writeup by EvilMushroom666
Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD
Cakes can and will pin! - So you think cakes suck for pins. Your wrong
Franks Simple Coir/Verm Tek
Franks Proper Pasturisation Tek
Franks Spawning To Bulk - Monotub
Professor Pinheads RTV Injection Port Tek
Foo Mans No Soak WBS Prep Tek


Edited by veda_sticks (12/07/07 05:42 AM)


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OfflineSaryn
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Re: Fighting contams, could use some advice [Re: veda_sticks]
    #7747770 - 12/12/07 12:33 AM (16 years, 1 month ago)

Just wanted to post a quick update. Looks like the contamination was coming from moving air in the closet I was using to incubate the jars. Simply added some aluminum foil over the entire top of the lid (not air tight) to keep things from settling in the dry verm. Have not had one of the jars still colonizing show up with tich since. The only way I can figure that the tich was then making it to the wet verm was when I would rotate the jars to watch the growth progress, it shook the tich down.

Will end up having a merry xmas after all! hehe


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Invisibletahoe
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Re: Fighting contams, could use some advice [Re: Saryn]
    #7747888 - 12/12/07 01:54 AM (16 years, 1 month ago)

there was a thread in the contam forum just the other day where another guy got a syringe from spores101 and he was getting some gren mold at teh injection sites.seems like someone is putting out dirty syringes


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


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Invisibletahoe
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Re: Fighting contams, could use some advice [Re: tahoe]
    #7747892 - 12/12/07 01:58 AM (16 years, 1 month ago)



--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


My Legacy
https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987

Teh=The
I need to proofread


Edited by tahoe (12/12/07 02:50 PM)


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OfflineSubstance D
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Re: Fighting contams, could use some advice [Re: tahoe]
    #7748415 - 12/12/07 08:55 AM (16 years, 1 month ago)

Quote:

tahoe said:
http://www.shroomery.org/forums/showflat.php/Number/7648341#7648341


i didnt see them mention spores101 in that thread,i hope there neddles are good as i just got 2 in the mail


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OfflineRogerRabbitM
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Re: Fighting contams, could use some advice [Re: Substance D]
    #7748445 - 12/12/07 09:08 AM (16 years, 1 month ago)

Don't forget to flame sterilize the needle until it's red hot between each jar. Wipe down the micropore tape with alcohol before injecting through it. This ensures you're not cross-contaminating between jars.

I doubt seriously that air movement in your closet is to blame. I colonize jars on an open shelf in my bedroom, and they get plenty of moving, dirty air around them all the time.

Make sure you're using fine vermiculite for the filter. Course will not stop the mold spores as well. using coffee filters over your jars to catch contaminants would be like trying to catch a gnat in a butterfly net. They won't help. If you want to keep dust/mold spores off your jars, wrap a piece of tyvek over the top. Good luck.
RR


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OfflineSpores_101
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Re: Fighting contams, could use some advice [Re: RogerRabbit]
    #7748694 - 12/12/07 10:46 AM (16 years, 1 month ago)

I didnt see any mention of us in that thread either..please don't sling shit if the info isnt there.  :shitstorm:


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Invisibletahoe
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Re: Fighting contams, could use some advice [Re: Spores_101]
    #7749734 - 12/12/07 02:51 PM (16 years, 1 month ago)

sorry didnt mean to link the wrong thread. It is here
http://www.shroomery.org/forums/showflat.php/Number/7741529#7741529


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


My Legacy
https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987

Teh=The
I need to proofread


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InvisibleGretchenmeister
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Re: Fighting contams, could use some advice [Re: RogerRabbit]
    #7750631 - 12/12/07 06:27 PM (16 years, 1 month ago)

I like a verm layer, and I always got contams if i didnt cover the innoc holes. Also, be sure you are using an aerosol with quatenary ammonium and not pentols. (lysol and oust have pentols, clorox aerosol has quat ammonium). Additionally, I never thought it hurt to completley dip the sryinge in peroxide btwn injections...I have gone around with contams before and found i needed to be sterilizing the needle closer to where it meets the plastic...without melting it.
I have also been known to flame sterilize the injection sites. I have always liked spores101. Hope that helps some.


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