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damn trich is back


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#1 teletrue

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Posted 24 September 2006 - 04:44 PM

About a month ago I posted how, despite my best OCD efforts, I got trich in every jar I noc'd up...I've never dealt with trich like this before. This is horrible. I can't get one jar to come up clean for me.

I've always steamed my jars the same way, and never enountered anything like this. I've had one jar that came up with contamination, and the fungus was black, and only 2-3 jars, out of...60? that have contained trich. And those only came up when I wasn't being clean. I really can't pinpoint what's doing it. I showed pics last time of how the BRF/Verm in the jar turned sort of a greenish color (like someone had shot green dye in the jar), and then after this was exposed to light and heat for a few days, this is what came up...

Dammit.

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#2 Sky Rayven

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Posted 24 September 2006 - 04:54 PM

Are you steaming or are you pcing?

Maybe try using a glove box.

Or try shooting up at another location??

Check your pad for mold and mildew.

Shit Shower and Shave before hand.

I really dunno...these are my best suggestions.

#3 Lazlo

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Posted 24 September 2006 - 05:02 PM

That mold appears to be comming from the inoculation, not the substrate.

#4 teletrue

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Posted 24 September 2006 - 05:05 PM

That mold appears to be comming from the inoculation, not the substrate.


Well, I got the spores from a reliable vendor here on mycotopia..

*edit* I've always used spores from a different vendor. This is the first time I've tried this vendors spores...

#5 apokalypse

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Posted 24 September 2006 - 05:23 PM

Could just be a contaminated syringe. Do you have any spare prints? Clean out the syringe with boiling hot water, and make up a syringe. Or get in touch with that vendor and see what they can do for you.

#6 srgtm1a

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Posted 24 September 2006 - 05:24 PM

Fractional Sterilization does work, but as you can see, isn't always sure fire.

Possibly look into getting a cheap PC, and look closer at your sterilization techniques. Not only for the jars, but for syringe sterilization as well.

Sometimes it is the smallest things we overlook that cause the green meanie.

Although vendors do sometimes get contaminated syringes, I don't think that is the case this time.

PS: If you are doing 60 BRF jars....you need to look into bulk growing and other methods of cultivation.

#7 apokalypse

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Posted 24 September 2006 - 05:29 PM

srgtm1a, see how there is that pukey looking light greenish mold right in the center of the trich? I've had that happen before after innoculation, using a re-used and cleaned syringe. But that was also from steaming jars for an hour, so you could be on to something.

#8 srgtm1a

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Posted 24 September 2006 - 05:34 PM

I think the mold is from the inoculation. I agree with lazlo on that. It's fairly obvious.

But I don't think it is a problem from the vendor's syringe. I think this is due to a mix of fractional sterilization and ill prep of the syringe b4 inoculation.

That part in the center, I'm willing to bet, is where the spore solution was sprayed.

It could be a ton of different reasons it happened, Contaminated syringe, too much spore solution added to substrate, lack of good syringe sterilization....so on and so on. In a situation where it keeps happening, and happens through multiple vendor syringes, you really need to hunker down, and go over sterilization step by step.

I re-use syringes all the time. So long as you take the proper steps in sterilization, it's not a problem.

#9 I_am_me

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Posted 24 September 2006 - 05:42 PM

He isn't doing fractional sterilization. That is where you try to sterilize grain jars 3 times or so in 3 days, allowing endospores to germinate and then killing them off. He is just sterilizing with steam. He has pf tek jars right? Steaming those jars is just fine and I've done so with a very low contam rate, maybe 1 to 2 jars per 40 or so. Just to be sure though you could try steaming them for an extra 30 minutes but generally the temperatures reached through steaming are enough to properly sterilize pf jars.

Did you prepare the syringes yourself from a print from a vendor or did you just buy premade syringes? It looks to me like it wasn't from just the innoculation point unless a whole lot of spore solution was added. Usually the first growth from an innoculation point will occur at the center. I had jars contam in the pattern shown in the jar above but it was due to bad air getting in through the verm barrier.

Hehe and yes, while to some preparing and fruiting 60 pf jars is a lot some people still prefer it over having to deal with the issues that accompany bulk substrates. ;) Then again my motto is variety is the spice of life, use all the teks. :D

#10 teletrue

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Posted 24 September 2006 - 05:44 PM

Well, this all started over the summer, when I got my first initial shipment of syringes (3 to be exact). The jars that were noc'd up with the EQ LC all came up green (12 jars)..after a post on here where some, including moderators, thought it was a bad syringe, mine was replaced free of charge.

So I got the new EQ syringe, and noc'd up several jars recently with it, and the two other syringes. Everything was done as I have always done it, and every single jar came up with Trich.

I think I'm going to get a syringe from my old vendor and see what happens. Just for shits and giggles.

#11 I_am_me

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Posted 24 September 2006 - 05:48 PM

I at least think the problem isn't your sterilization techniques. Maybe if you get a new syringes try to document the whole process well so if it still works out poorly then we can have some more info to diagnose the problem. Best of luck.

#12 srgtm1a

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Posted 24 September 2006 - 05:52 PM

Any kind of steam sterilization is considered Fractional Sterilization. The use of doing it with grain is a technique using fractional sterilization over a number of days to prep grain, which, IMO does not work with grain..

The syringe he is using is from a vendor, not self made. The contam he is seeing is not caused by "bad air" getting in, it is directly caused by the inoculation. His pictures are identical to every picture i've ever seen of contaminated syringes and or improper steriliztion b4 inoculation.

The reason I know it is a problem with his sterilization techniques is because this has happend to him multiple times, using multiple vendors. Chance of a vendor replacing a contaminated syringe with another contaminated syringe are slim to none.

#13 teletrue

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Posted 24 September 2006 - 05:55 PM

He isn't doing fractional sterilization. That is where you try to sterilize grain jars 3 times or so in 3 days, allowing endospores to germinate and then killing them off. He is just sterilizing with steam. He has pf tek jars right? Steaming those jars is just fine and I've done so with a very low contam rate, maybe 1 to 2 jars per 40 or so. Just to be sure though you could try steaming them for an extra 30 minutes but generally the temperatures reached through steaming are enough to properly sterilize pf jars.

Did you prepare the syringes yourself from a print from a vendor or did you just buy premade syringes? It looks to me like it wasn't from just the innoculation point unless a whole lot of spore solution was added. Usually the first growth from an innoculation point will occur at the center. I had jars contam in the pattern shown in the jar above but it was due to bad air getting in through the verm barrier.

Hehe and yes, while to some preparing and fruiting 60 pf jars is a lot some people still prefer it over having to deal with the issues that accompany bulk substrates. ;) Then again my motto is variety is the spice of life, use all the teks. :D


I prepare my jars by filling them with the substrate, foil lids, and putting them in a pot with a dishrag on the bottom with water about halfway up the jars, and when the water starts to boil, I adjust the temp. to keep it constant, cover the pot tightly with Al. Foil, and let it go for an hour and a half.

It says above the syringes came from a sponsor of this site. I have a couple prints that I won in a contest on here, and I'd like to make some syringes out of them (hillbilly and B+), but I don't know where to begin or which tek to follow as I've always used vendor's syringes.

And again, I've been doing this a while...using only steam, and have never had anything this bad. My gut's telling me that it's the syringes.

#14 I_am_me

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Posted 24 September 2006 - 05:57 PM

I always though use of the work fractional implied steaming more than once, sterilizing a fraction of the endospores at a time. I really don't think it means the same thing as steaming. Steaming them once is simply sterilization, no "fractional" needed as its all done at once time. I know for a fact that steaming is perfectly fine for pf jars. I and many other even get away with doing 45 minutes as opposed to a whole hour.

If the problem was really with the sterilization of the jars it wouldn't appear in that pattern. It is either from a dirty needle, dirty spore solution or a verm barrier that is too loose so unfiltered air is allowed in during/after innoculation. Also I think his problem with this much contams has come from only one vendor with one strain, correct? Usually syringes are made in batches so I would assume it is quite possible both eq syringes came from the same bad batch, maybe even the same print or batch of prints.

#15 teletrue

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Posted 24 September 2006 - 05:57 PM

Any kind of steam sterilization is considered Fractional Sterilization. The use of doing it with grain is a technique using fractional sterilization over a number of days to prep grain, which, IMO does not work with grain..
The syringe he is using is from a vendor, not self made. The contam he is seeing is not caused by "bad air" getting in, it is directly caused by the inoculation. His pictures are identical to every picture i've ever seen of contaminated syringes and or improper steriliztion b4 inoculation.
The reason I know it is a problem with his sterilization techniques is because this has happend to him multiple times, using multiple vendors. Chance of a vendor replacing a contaminated syringe with another contaminated syringe are slim to none.


FYI, the last batch I did (40 PF jars, all steam sterilized, and innoculated using a glove box last year) came out clean and produced gobs of shrooms... The contamination I was referring to was when I first started this hobby.

#16 srgtm1a

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Posted 24 September 2006 - 06:02 PM

I always though use of the work fractional implied steaming more than once, sterilizing a fraction of the endospores at a time. I really don't think it means the same thing as steaming. Steaming them once is simply sterilization, no "fractional" needed as its all done at once time. I know for a fact that steaming is perfectly fine for pf jars. I and many other even get away with doing 45 minutes as opposed to a whole hour.

If the problem was really with the sterilization of the jars it wouldn't appear in that pattern. It is either from a dirty needle, dirty spore solution or a verm barrier that is too loose so unfiltered air is allowed in during/after innoculation.


Fractional is in reference to pressure, not time.

I'm not saying there is anything wrong with steam or fractional sterilization. As I said....it does work, but isn't the best.

I agree that the source of the problem is at the inoculation, it's obvious just by the pattern of the mold, but I don't agree that it is a problem with the vendor's syringes.

This has happend to him before with a previous syringe. The chance that the free replacement also is contaminated is very unlikely. If that was the case, there would be a LOT more people crying foul.

Teletrue, go ahead and get a syringe from your old vendor, and if it still happens to you, then you know where the problem lies.

#17 teletrue

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Posted 24 September 2006 - 06:04 PM

Ok.

I've decided that tonight i'm going to make up a few jars and noc them up tomorrow.

I plan on steaming them in a tightly covered pot for an hour and a half.

Give me advice here, tell me what to do.

*edit* I'm also going to photodocument everything and post the pictures here showing my procedure

#18 I_am_me

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Posted 24 September 2006 - 06:04 PM

http://mycotopia.net...isplay.php?f=59

There are several easy teks for syringe making there. The only thing I would recommend is making a glovebox to do your syringe making in. I don't need one myself but I live in a very cold part of the country and have little trouble with contams all together. A glovebox will really help cutback on the troubles you would encounter. Other than that, it is all very easy if you read over the tek plenty before attempting. I even recommend doing a dry run without using a real print just to see if you run into any problems with your technique/procedures.

#19 I_am_me

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Posted 24 September 2006 - 06:07 PM

Fractional is in reference to pressure, not time.

I'm not saying there is anything wrong with steam or fractional sterilization. As I said....it does work, but isn't the best.

I agree that the source of the problem is at the inoculation, it's obvious just by the pattern of the mold, but I don't agree that it is a problem with the vendor's syringes.

This has happend to him before with a previous syringe. The chance that the free replacement also is contaminated is very unlikely. If that was the case, there would be a LOT more people crying foul.


http://www.biology-o...l_sterilization

Also I've found several other sources will all indication that fractional sterilization specifically refers to several runs through sterilization process with a day of incubation inbetween each, also referred to as tyndallization.

Also in the time I've been dealing with spore vendors I've seen more than once occasion that a bad batch of syringes was made. Many many syringes get replaced all the time and there are so many reasons that go into contamination that it isn't always practical to blame the vendor, even if that was the real issue. When you make as many syringes as a vendor does you're bound to have a bad batch every now and then. All it takes is one dirty print to make quite a few dirty syringes.

#20 srgtm1a

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Posted 24 September 2006 - 06:21 PM

http://www.biology-o...l_sterilization

Also I've found several other sources will all indication that fractional sterilization specifically refers to several runs through sterilization process with a day of incubation inbetween each, also referred to as tyndallization.

Also in the time I've been dealing with spore vendors I've seen more than once occasion that a bad batch of syringes was made. Many many syringes get replaced all the time and there are so many reasons that go into contamination that it isn't always practical to blame the vendor, even if that was the real issue. When you make as many syringes as a vendor does you're bound to have a bad batch every now and then. All it takes is one dirty print to make quite a few dirty syringes.


I've seen that from a few vendors as well, but in cases where it was a bad batch, there were a ton of complaints, and the whole batch was pulled and replaced at later dates. The last vendor I saw this happen to was spores101, only they never admitted to it.

Reading his past problems etc, leads me to belive it is a problem with his syringe sterilization, and not the vendor. Sure it happens to vendors from time to time, but not often enough to be likely in this case.

As far as fractional sterilization goes, every chemistry class i've ever taken in college refered to it as sterilization at a fraction of the pressure used in a lab environment, done once, not at multiple intervals.




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