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Invisiblebootster
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Why does this happen to all my jars?
    #19328290 - 12/26/13 10:40 AM (10 years, 1 month ago)

I don't have a clue.



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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328295 - 12/26/13 10:43 AM (10 years, 1 month ago)

Have you left any jars not inoculated to see whether it's a bunk syringe/sterile technique or your grain prep?


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19328312 - 12/26/13 10:48 AM (10 years, 1 month ago)

I always do that and they remain clean. I have 18 jars of different strains/species and it always shows up at once and soon they will have a powdery look to them. After a shake, the green comes out and forms a green cloud in the jar. Some of these are directly injected by a syringe from a vendor with a red hot needle into an RTV injection port. :shrug:

Most come from clean looking agar cultures.


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Re: Why does this happen to all my jars? [Re: bootster]
    #19328319 - 12/26/13 10:53 AM (10 years, 1 month ago)

maybe change the substrate supplier????


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InvisibleJuicin
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328326 - 12/26/13 10:55 AM (10 years, 1 month ago)

If you're using clean wedges i'd try changing spawn supplier...........

If you can do agar work in your house but can't get grains going that's weird as fuck. And i'd start by getting new grains


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InvisiblePirateSwazey
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328327 - 12/26/13 10:55 AM (10 years, 1 month ago)

you sure your agar has mushroom myc and not a similar lookin mold myc growing on it?

are you inoculating your dishes with clone tissue or spores?

I usually only catch bacteria on my petris but I have inoculated a jar with what
looked like a wedge of mushroom myc and turned out to be the green. it grew
with a ghost or grey looking myc on the grain though... not what yours looks like.


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: Mrdbrewer]
    #19328332 - 12/26/13 10:56 AM (10 years, 1 month ago)

Quote:

Mrdbrewer said:
maybe change the substrate supplier????




I grew them out clean on agar. What would changing a supplier accomplish?

No disrespect intended but some of these have been grown out 5 or 6 times.


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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328342 - 12/26/13 10:58 AM (10 years, 1 month ago)

Quote:

bootster said:
I always do that and they remain clean. I have 18 jars of different strains/species and it always shows up at once and soon they will have a powdery look to them. After a shake, the green comes out and forms a green cloud in the jar. Some of these are directly injected by a syringe from a vendor with a red hot needle into an RTV injection port. :shrug:

Most come from clean looking agar cultures.




I've had vendor syringes that contaminated everything with mold, but if you're getting green also when you inoculate with agar wedges and the jars you leave closed after the PC don't get mold in them, then it's clearly something to do with your sterile technique.  Do you use an SAB or a flow hood?


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19328418 - 12/26/13 11:19 AM (10 years, 1 month ago)

I am looking at clean mycelium.





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InvisiblePirateSwazey
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328433 - 12/26/13 11:23 AM (10 years, 1 month ago)

Maybe it's like these guys are saying...

something in your grain :shrug:

Have you tried pc'ing some WBS n using that since you've been having problems?


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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: PirateSwazey]
    #19328464 - 12/26/13 11:35 AM (10 years, 1 month ago)

Quote:

PirateSwazey said:
Maybe it's like these guys are saying...

something in your grain :shrug:

Have you tried pc'ing some WBS n using that since you've been having problems?




If you can work agar without contams it's not your sterile technique, are you sure you've tried leaving a few sterilized jars that have been PC'd without innoculating them to test to see if they come out properly sterilized?  Are you soaking 24 hours, are you adding gypsum?  How long do you PC?  Switch to a different source for grain, give it a good long soak, don't add any supplements at first and give it a good solid 90 minutes at 15 PSI.  I think it's your grain even though you claim to leave test jars.  Might be your gypsum as I've had mold like that the few times I left a bit too much residual gypsum on the grains before PC-ing them.  :shrug:

If it's not your grain then go on to the next step, which is finding out where you're fucking up your sterile technique.


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328478 - 12/26/13 11:41 AM (10 years, 1 month ago)

I just called my supplier and he said that the rye was absolutely not treated.


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Invisible36fuckin5
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Re: Why does this happen to all my jars? [Re: PirateSwazey]
    #19328479 - 12/26/13 11:41 AM (10 years, 1 month ago)

What kind of filter are you using? What are your procedures for sterilization? How confident are you in transferring agar to grain? It's not hard, but it is kinda tricky to learn to do it just right.

I doubt it's your grain source. That just sounds kind of retarded. All grain is grown outside, stored in a silo or barn and is covered in all kinds of contams.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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InvisibleJuicin
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Re: Why does this happen to all my jars? [Re: 36fuckin5]
    #19328534 - 12/26/13 11:55 AM (10 years, 1 month ago)

Grain supply being tainted for whatever reason makes a lot more sense than a 100% failure rate from agar to grain.


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328538 - 12/26/13 11:56 AM (10 years, 1 month ago)



I've never had any problems before and it's been going on for a long time now. I am spending way too much $$$ just to see fruits.

I use a SAB of my own design. I've done many monotubs before. I've grown pounds of Shiitake, Blue Oyster, King Oyster, Lions Mane and many other kinds of edibles using two separate suppliers. I've never had a problem.
I'm using Prof P's lid TEK (before he started to demean me) with a 3/8" hole stuffed with poly fill and aluminum caps.

I'm thinking that the grain is too dry. I went on a bizarre run for a while when the grains were too wet and had tons of bacteria issues.

I just don't get it, unless the Tyvek sleeves in my SAB bring contams into it. :shrug:


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Invisible36fuckin5
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328558 - 12/26/13 12:04 PM (10 years, 1 month ago)

Your filter is probably your problem. I'm having the same issue. Polyfil just isn't that great unless your environment is fairly clean.

Get some SFDs from Out-Grow or Aloha (the stupidly thick ones, like 1/16"), use them whole as intended and check your house thoroughly for mold.

You also didn't answer my question about your sterilization procedure. I need every detail, too.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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OfflineAero
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Re: Why does this happen to all my jars? [Re: 36fuckin5]
    #19328584 - 12/26/13 12:12 PM (10 years, 1 month ago)

might be a problem in the sterilization process
not enough pressure in the PC?

i would do a few verm+whole brown rice jars to see whats the problem

2part verm 1 part rice 1part water


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Invisible36fuckin5
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Re: Why does this happen to all my jars? [Re: Aero]
    #19328590 - 12/26/13 12:14 PM (10 years, 1 month ago)

Quote:

Aero said:
i would do a few verm+whole brown rice jars to see whats the problem

2part verm 1 part rice 1part water




WTF would this teach you about this particular problem?


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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OfflineAero
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Re: Why does this happen to all my jars? [Re: 36fuckin5]
    #19328598 - 12/26/13 12:16 PM (10 years, 1 month ago)

Quote:

36fuckin5 said:
Quote:

Aero said:
i would do a few verm+whole brown rice jars to see whats the problem

2part verm 1 part rice 1part water




WTF would this teach you about this particular problem?





that the culture+syringe+environment is clean my friend, and the problem is the grain

think before u start raging


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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: Aero]
    #19328605 - 12/26/13 12:19 PM (10 years, 1 month ago)

Checking the filters is a good idea.  What tek are you using for your grain jar lids?


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OfflinecronicrFacebook
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Re: Why does this happen to all my jars? [Re: Aero]
    #19328607 - 12/26/13 12:19 PM (10 years, 1 month ago)

Quote:

Aero said:
Quote:

36fuckin5 said:
Quote:

Aero said:
i would do a few verm+whole brown rice jars to see whats the problem

2part verm 1 part rice 1part water




WTF would this teach you about this particular problem?





that the culture+syringe+environment is clean my friend, and the problem is the grain

think before u start raging



Agar would be the right suggestion, cakes can be steamed so the answer wouldn't help much, be sure to vent your steam for a few minutes before putting your weight on


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OfflineAero
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Re: Why does this happen to all my jars? [Re: cronicr]
    #19328618 - 12/26/13 12:23 PM (10 years, 1 month ago)

Quote:

cronicr said:
Quote:

Aero said:
Quote:

36fuckin5 said:
Quote:

Aero said:
i would do a few verm+whole brown rice jars to see whats the problem

2part verm 1 part rice 1part water




WTF would this teach you about this particular problem?





that the culture+syringe+environment is clean my friend, and the problem is the grain

think before u start raging



Agar would be the right suggestion, cakes can be steamed so the answer wouldn't help much, be sure to vent your steam for a few minutes before putting your weight on




he is using agar


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: Aero]
    #19328628 - 12/26/13 12:26 PM (10 years, 1 month ago)

I started out with monotubs and never have had an interest in cakes. I have had nothing but good luck with monotubs and it is frustrating to have everything go by the wayside.

Sterile techniques are the same as they ever were (without going into detail :shrug: ).

I am thinking about going with plastic lids and SFD patches siliconed on.

I am lazy and when a jar sits with green in it, it actually stains the SFD green. I don't know if I'm comfortable with reusing them. The plastic idea may not work either.

Thank you guys for taking time to answer my questions and try to help me out.


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Invisible36fuckin5
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Re: Why does this happen to all my jars? [Re: Aero]
    #19328641 - 12/26/13 12:32 PM (10 years, 1 month ago)

Quote:

Aero said:
that the culture+syringe+environment is clean my friend, and the problem is the grain

think before u start raging




The problem is obviously either inoculant, technique or bad filters.

If you're venting your PC for a few minutes and running a full 15 PSI for 90 minutes, the grain is fine.

The agar plates look clean, so it's not inoculant, because it happens with vendor syringes and agar wedges. He can apparently get clean plates, so I'd imagine he can probably toss one in a grain jar without contaminating it, since it's the same motions.

Therefore, I'm blaming the filters. Polyfil isn't the best thing in the world to use. It can get jostled and open holes, and it also just isn't the best pore size for filtration without many, many layers, and just how many you need hasn't really been established.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: bootster]
    #19328651 - 12/26/13 12:33 PM (10 years, 1 month ago)

Quote:

bootster said:
I started out with monotubs and never have had an interest in cakes. I have had nothing but good luck with monotubs and it is frustrating to have everything go by the wayside.

Sterile techniques are the same as they ever were (without going into detail :shrug: ).

I am thinking about going with plastic lids and SFD patches siliconed on.

I am lazy and when a jar sits with green in it, it actually stains the SFD green. I don't know if I'm comfortable with reusing them. The plastic idea may not work either.

Thank you guys for taking time to answer my questions and try to help me out.




Lids with SFD glued on are the way to go, hopefully you can just avoid getting the green in your jars in the first place and you won't have to worry about the mold ruining your filters for future use.  :lmafo:


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Invisible36fuckin5
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19328723 - 12/26/13 12:49 PM (10 years, 1 month ago)

Just don't get those thin-ass cheap (they're not even inexpensive, just cheap) SFDs. Get the super thick ones from Out-Grow. And I wouldn't try cutting them, just use them hole. If you have to use a syringe one day, just throw it in your SAB, pop the lid and squirt some in.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19328787 - 12/26/13 12:59 PM (10 years, 1 month ago)

Quote:

RiparianZoneJunky said:
Quote:

bootster said:
I started out with monotubs and never have had an interest in cakes. I have had nothing but good luck with monotubs and it is frustrating to have everything go by the wayside.

Sterile techniques are the same as they ever were (without going into detail :shrug: ).

I am thinking about going with plastic lids and SFD patches siliconed on.

I am lazy and when a jar sits with green in it, it actually stains the SFD green. I don't know if I'm comfortable with reusing them. The plastic idea may not work either.

Thank you guys for taking time to answer my questions and try to help me out.




Lids with SFD glued on are the way to go, hopefully you can just avoid getting the green in your jars in the first place and you won't have to worry about the mold ruining your filters for future use.  :lmafo:




I'm not laughing but you have made a good point. I think I'll pursue that avenue for failure.

Thanks guys.

Here is a load of jars that I did last night.








Edited by bootster (12/26/13 01:20 PM)


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OfflineKizzle
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Re: Why does this happen to all my jars? [Re: 36fuckin5]
    #19328829 - 12/26/13 01:08 PM (10 years, 1 month ago)

A culture can be contaminated without showing any signs. That scenario usually happens though as a result of the culture becoming contaminated after it is already at least partially colonized, so it'd still most likely be related to your sterile technique.

I could see it possibly being the filter if you treated your control jars differently than the incoulated jar, like if you didn't shake them but shook the incoulated ones or stored them in a different location.


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Invisiblebootster
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Re: Why does this happen to all my jars? [Re: Kizzle]
    #19328864 - 12/26/13 01:23 PM (10 years, 1 month ago)

You may have a point Kizzle but I usually transfer up to 7 or 8 times (I'm anal about transfers) before I'm confident that the culture "looks" right.


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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: bootster]
    #19329129 - 12/26/13 02:27 PM (10 years, 1 month ago)

Quote:

bootster said:
You may have a point Kizzle but I usually transfer up to 7 or 8 times (I'm anal about transfers) before I'm confident that the culture "looks" right.




When I've had cultures get mold from neglect the contam shows up on the next plate after a transfer, so if you're doing multiple transfers and don't see contams on agar it's definitely not your agar wedges.  The fact that you work with agar and aren't getting contams on your plates shows that you have good sterile technique in general and making transfers is hardly different in terms of technique than throwing a wedge in a grain jar.  It's gotta be your polyfil.  I just cut small discs of SFD and silicone glue them over your holes, you don't even need an injection port if you're going to be exclusively noccing with agar.  I use this tek and have zero problems.  Good luck :grin:


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OfflineKizzle
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19329162 - 12/26/13 02:36 PM (10 years, 1 month ago)

There's nothing wrong with polyfill in general but if it's getting wet that could be a problem.


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OfflineAero
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Re: Why does this happen to all my jars? [Re: Kizzle]
    #19329536 - 12/26/13 04:26 PM (10 years, 1 month ago)

yea i too use polyfil with zero contam rate..


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Re: Why does this happen to all my jars? [Re: Aero]
    #19329556 - 12/26/13 04:31 PM (10 years, 1 month ago)

I know it's hard to tell but those grains don't look hydrated to me. I'd make sure your pressure cooker is maintaining the correct temperature. Remember, agar will take a lot less time to sterilize then grain.


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Re: Why does this happen to all my jars? [Re: jellyfish]
    #19329574 - 12/26/13 04:35 PM (10 years, 1 month ago)

Quote:

jellyfish said:
I know it's hard to tell but those grains don't look hydrated to me. I'd make sure your pressure cooker is maintaining the correct temperature. Remember, agar will take a lot less time to sterilize then grain.




That is a good point. I don't think anyone else has offered the PC being an issue here :thumbup:


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OfflineAero
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Re: Why does this happen to all my jars? [Re: PirateSwazey]
    #19329687 - 12/26/13 05:02 PM (10 years, 1 month ago)

Quote:

PirateSwazey said:
Quote:

jellyfish said:
I know it's hard to tell but those grains don't look hydrated to me. I'd make sure your pressure cooker is maintaining the correct temperature. Remember, agar will take a lot less time to sterilize then grain.




That is a good point. I don't think anyone else has offered the PC being an issue here :thumbup:




i was actually

Quote:

Aero said:
might be a problem in the sterilization process
not enough pressure in the PC?

i would do a few verm+whole brown rice jars to see whats the problem

2part verm 1 part rice 1part water




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SPREAD THE SPORES



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Re: Why does this happen to all my jars? [Re: Aero]
    #19329706 - 12/26/13 05:07 PM (10 years, 1 month ago)

so did i, but i do believe your right about the grains, they look like they didn't get to soak enough


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I'm tired do me a favor


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Re: Why does this happen to all my jars? [Re: cronicr]
    #19329731 - 12/26/13 05:13 PM (10 years, 1 month ago)

^ Agreed.

I think OP said he was trying to keep them from soaking too long to prevent bacteria.
Could be possible that by not soaking long enough he isn't germinating enough endospores :popcorn:

Sorry I must have missed all the other input concerning the PC...


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OfflinecronicrFacebook
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Re: Why does this happen to all my jars? [Re: PirateSwazey]
    #19329733 - 12/26/13 05:14 PM (10 years, 1 month ago)

i think it would be more along the lines of it being harder for the pressure to penetrate but i could be wrong


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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19329760 - 12/26/13 05:23 PM (10 years, 1 month ago)

Quote:

PirateSwazey said:
^ Agreed.

I think OP said he was trying to keep them from soaking too long to prevent bacteria.
Could be possible that by not soaking long enough he isn't germinating enough endospores :popcorn:

Sorry I must have missed all the other input concerning the PC...




Quote:

RiparianZoneJunky said:
Quote:

PirateSwazey said:
Maybe it's like these guys are saying...

something in your grain :shrug:

Have you tried pc'ing some WBS n using that since you've been having problems?




Switch to a different source for grain, give it a good long soak, don't add any supplements at first and give it a good solid 90 minutes at 15 PSI.  I think it's your grain even though you claim to leave test jars. 





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Edited by RiparianZoneJunky (12/26/13 05:24 PM)


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OfflinecronicrFacebook
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19329762 - 12/26/13 05:24 PM (10 years, 1 month ago)

as long as he hydrates right the grain choice won't matter
When the grains are hydrated, the hard shell of the endospore is also hydrated and will be killed in the pc weather or not it germinated

oops that was a rr qoute i just forgot to qoute instead i just copied


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It doesn't matter what i think of you...all that matters is clean spawn

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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: cronicr]
    #19329769 - 12/26/13 05:26 PM (10 years, 1 month ago)

Quote:

cronicr said:
as long as he hydrates right the grain choice won't matter
When the grains are hydrated, the hard shell of the endospore is also hydrated and will be killed in the pc weather or not it germinated



All I'm saying is the first thing I told OP was to make sure his grain prep is solid.    :shrug:


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OfflineAero
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Re: Why does this happen to all my jars? [Re: cronicr]
    #19329849 - 12/26/13 05:50 PM (10 years, 1 month ago)

Quote:

cronicr said:
as long as he hydrates right the grain choice won't matter
When the grains are hydrated, the hard shell of the endospore is also hydrated and will be killed in the pc weather or not it germinated

oops that was a rr qoute i just forgot to qoute instead i just copied





:whathesaid:


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OfflineKizzle
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Re: Why does this happen to all my jars? [Re: PirateSwazey]
    #19330275 - 12/26/13 07:42 PM (10 years, 1 month ago)

Quote:

PirateSwazey said:
Quote:

jellyfish said:
I know it's hard to tell but those grains don't look hydrated to me. I'd make sure your pressure cooker is maintaining the correct temperature. Remember, agar will take a lot less time to sterilize then grain.




That is a good point. I don't think anyone else has offered the PC being an issue here :thumbup:



If it was bacteria perhaps since some are very heat resistant but it's not realistic for sterilization to be the cause of mold in a spawn jar.

It wouldn't be the grain prep either. Granted some molds can only grow on damaged grains in any case it would still mean mold is getting into the jar somehow after sterilization.


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Re: Why does this happen to all my jars? [Re: Kizzle]
    #19330634 - 12/26/13 09:09 PM (10 years, 1 month ago)

you're the contam major kizzle not me :wink:

I thought it was bacterial


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OfflineChaostoOrder
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Re: Why does this happen to all my jars? [Re: 36fuckin5]
    #19330659 - 12/26/13 09:15 PM (10 years, 1 month ago)

For one your rye looks wayyy to dry.  Boiling them after a 12 hour soak is a good way to add water to the rye. Its not necessarily a way to boil out bacteria, because we all know you will never fully rid them of that...but yeah you should boil your rye after your soak.  That mycellium looks like its roughin it bro... is that some trich I see above your mycellium????


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InvisibleHypnotoadCroaked
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Re: Why does this happen to all my jars? [Re: ChaostoOrder]
    #19330685 - 12/26/13 09:21 PM (10 years, 1 month ago)

Contamination need to be introduced to sterile media.  If a "test" jar is left uninoculated and does not contaminate, than it must be the method of introducing a culture OR the culture itself. 

The water density of sterile media has no impact on mold growing.  If mold is growing, I would deduce that water content is close to correct.

If this is happening EVERY time, it is time to clean house.  Get new prints and start from square one.  It can happen.


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OfflineRiparianZoneJunky
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Re: Why does this happen to all my jars? [Re: HypnotoadCroaked]
    #19330754 - 12/26/13 09:42 PM (10 years, 1 month ago)

Let us know what you end up doing and how it works out OP, seems to me you have a wealth of options to try. :lol:  I still think it's either your grain prep or your filter, I do too much agar to buy the idea that there are hidden contams on your cultures that only grow when you put them on grain, you clearly have enough sense to make a transfer without growing green on agar and that's way harder than noccing jars.  :shrug:


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Offlinetwistedty
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Re: Why does this happen to all my jars? [Re: RiparianZoneJunky]
    #19331358 - 12/27/13 12:58 AM (10 years, 1 month ago)

try a better filter and report back after using your same cultures


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