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 factory81
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 WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics 
#11042898 - 09/11/09 07:57 PM (2 years, 8 months ago) |
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Okay,
I had a (2) B+ syringes from ralph. I inoculate 12 jars that I was reusing. Couple days later we have contamination.
I have half a syringe of the B+ left. I inoculate 4 more brand new jars thinking well maybe the lids were contaminated. Same problem. So we are done with the B+ syringes which resulted in 100% failure of every jar.
Now we are on to the next try...
We inoculate 12 more jars and have 100% failure. This is with brand new jars, cleaned before filled. 90 minutes of steralizing. Remove from heat source. Let the jars cool for 12-18 hours. Inoculate the jars. In between every couple jars I would take a lighter to the syringe to try and steralize it.
We used gloves, a breathing mask, and we thoroughly clean the area in which we make the jars, and inoculate the jars.
I am confused, and something about me thinks its the syringes I've been getting from Ralph because this has worked time and time again with no problems. And then all of a sudden we have 100% contamination rate.
The contamination begins at the injection site and spreads.
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lsatrap
Mush maker
 
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Re: WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics [Re: factory81]
#11043399 - 09/11/09 09:38 PM (2 years, 8 months ago) |
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Yea I would say that your syringe is contaminated.Contaminates in the needle are very hard to kill even when flamed. We usually flame the needle to kill contames outside of the needle. Just a thought.
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dstark
Yes you can but you may Not!
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Re: WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics [Re: lsatrap]
#11045621 - 09/12/09 08:58 AM (2 years, 8 months ago) |
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Quote:
lsatrap said:
Yea I would say that your syringe is contaminated.Contaminates in the needle are very hard to kill even when flamed. We usually flame the needle to kill contames outside of the needle. Just a thought.
You have to flame it until it is red, that means the syringe temp is over 500 and contams have no chance.
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lsatrap
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Re: WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics [Re: dstark]
#11046318 - 09/12/09 11:39 AM (2 years, 8 months ago) |
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Quote:
dstark said:
Quote:
lsatrap said:
Yea I would say that your syringe is contaminated.Contaminates in the needle are very hard to kill even when flamed. We usually flame the needle to kill contames outside of the needle. Just a thought.
You have to flame it until it is red, that means the syringe temp is over 500 and contams have no chance.
For this to work you would have to flame the entire needle not just the end of it. I've learned this the hard way.Sometimes contams get stuck in the needle near the plastic part of the syringe so if you only flame the end of the needle you are only getting part of the contams killed.I'm not saying that the OP did this. I'm just saying it has happened to me..
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rorschachX
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Re: WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics [Re: factory81]
#11051116 - 09/13/09 09:23 AM (2 years, 8 months ago) |
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Quote:
I inoculate 12 jars that I was reusing. Couple days later we have contamination.
What method did you use to ensure the jars were clean and sterile prior to reusing? my FOAF prefer to sterilize them before he adds his grain and put into his AA sterilizer; this gives an N+1 (redundancy) situation which is not a bad thing.
Quote:
I inoculate 4 more brand new jars thinking well maybe the lids were contaminated. Same problem. So we are done with the B+ syringes which resulted in 100% failure of every jar.
While contam'd syringes CAN happen my FOAF finds it unlikely when ordering from a good source such as Ralphs or sporeworks. This is what they do for their business and additionally they always use fresh syringes and then place into a sterile sealed bag. I can not speak for Ralphs but Spore works does not even place the needle on your syringe; they leave it in the sterile wrapping for you to put on yourself.
Not saying it can't happen but my FOAF thinks it seems not to be the likely culprit.
Quote:
Now we are on to the next try...
We inoculate 12 more jars and have 100% failure. This is with brand new jars, cleaned before filled. 90 minutes of steralizing. Remove from heat source. Let the jars cool for 12-18 hours. Inoculate the jars. In between every couple jars I would take a lighter to the syringe to try and steralize it.
That's 26 jars now with a 100% contamination rate. What method did you use to clean your jars? What method did you use to sterilize? What equipment do you have to perform your sterilization? What method are you using for gas exchange? What is your method for injecting each jar with spores?
All of this may seem dumb but these are all important questions.
Did you use Bleach Water followed by soap/water to wash your jars? PC'ing for 30 minutes at 15-17psi followed by a wash would also be good.
Did you Sterilize in a pressure cooker? If so what brand? Does it have a gauge or does it have a rocker weight on it? Is a metal to metal seal or does it use a gasket? If you have a gasket type I would recommend replacing it, but if this is something you do regularly (you did 26 jars so my FOAF assumes so) my FOAF would recommend an All American since they have a gauge and a metal to metal seal. You could have one already but it didn't say in your post so I thought I'd point it out.
Are you using a metal Ring with Tyvek (that's what it looks like) or are you using something with polyfill? Or are you using a combination? How are you injecting into these jars? And if you're j ust injecting by puncturing through the tyvek/polyfill then what method are you using to make sure contamination doesn't enter through the injection point? Remember that whatever method you use has to be sterile also. For example: My FOAF uses quart jars with metal rings and tyvek replaced for the lids; he injects through the center of the tyvek and quickly places a bandaid over the injection point. Bandaids come sterile out of their wrapping.
Quote:
We used gloves, a breathing mask, and we thoroughly clean the area in which we make the jars, and inoculate the jars.
That is great. My FOAF uses a tyvek suit with hoodie (painters suit with hood) gloves, mask, goggles and sterile booties. In addition to this he uses a glove box that has been cleaned with bleach water prior to use and then wiped down with 91% alcohol and allowed to dry. He then places all of his jars (as many as he can fit) all of his syringes and tools (including bandaids, alcohol wipes etc etc) and sprays everything in that glove box with Oust for about 20 seconds straight. He places the lid on the box and comes back in 2-5 minutes.
At this point he starts flame sterilizing the needle until it is glowing red hot, wraps the needle in an alcohol soaked wipe and punctures through the top of the tyvek. Before he injects he places a bandaid on the lid near the needle. He undoes the bandaid so just the cotton part shows. He injects about 2cc of spores per jar; as he's removing the needle with one hand he uses the other hand to place the bandaid over the hole and makes sure the adhesive is secure to the tyvek. Bandaids are breathable and sterile. He then takes the jar out of the glove box and sets it aside.
Quote:
I am confused, and something about me thinks its the syringes I've been getting from Ralph because this has worked time and time again with no problems. And then all of a sudden we have 100% contamination rate.
The contamination begins at the injection site and spreads.
It could be the syringes but again I find that unlikely since Ralph probably practices better sterility than most people on the forum but it is a possibility.
What grain were you innoculating? It looks like cracked corn or something that had many seeds/kernels burst open during the simmering process.
In either case my FOAF knows that mold will cause condensation against the glass of a jar; however your jar does look a bit wet. Stale wet and warm conditions are great for mold. My FOAF always errs on the side of caution and blowdry's his grain after he's had a go at it with draining, going through it with a paper towel and then finally blow drying it on the cool setting.
This makes sure there is no external moisture. Also the fewer burst kernals you have the better. Burst seed/kernals don't retain moisture and it ends up creating excess moisture within the jar (but if you do the towel/blow dry method this isn't so much an issue anyway).
Again that moisture could be from mold but the grain looks suspicious and I'm curious what it is and if the extra moisture in grain and the extra starches from cracked/burst grain are effecting this.
What temperature are you allowing your jars to colonize at?
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lsatrap
Mush maker
Registered: 08/07/05
Posts: 1,072
Loc: S.E USA
Last seen: 10 hours, 41 minutes
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Re: WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics [Re: rorschachX]
#11054154 - 09/13/09 07:46 PM (2 years, 8 months ago) |
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I still think it looks as if the mold spores where inoculated into the substrate.It doesn't look as if it was already in the sub and then grew. It looks like it was injected in.It might not be the vendors fault but I think the contams where introduced through inoculation.
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rorschachX
Stranger
Registered: 08/16/09
Posts: 30
Last seen: 2 years, 7 months
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Re: WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics [Re: lsatrap]
#11054640 - 09/13/09 09:11 PM (2 years, 8 months ago) |
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Quote:
lsatrap said:
I still think it looks as if the mold spores where inoculated into the substrate.It doesn't look as if it was already in the sub and then grew. It looks like it was injected in.It might not be the vendors fault but I think the contams where introduced through inoculation.
Absolutely could be. Easy test would to set aside a grain jar after sterilizing it and don't inoculate it. It would just allow you to say for certain it is the spore syringe.
My FOAF always sets aside 1 grain jar each time he knocks up a set of jars just for this reason.
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lsatrap
Mush maker
Registered: 08/07/05
Posts: 1,072
Loc: S.E USA
Last seen: 10 hours, 41 minutes
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Re: WTF 3 tries - Inoculation = green/blue contam - Bad syringes from Ralph? pics [Re: rorschachX]
#11055223 - 09/13/09 10:31 PM (2 years, 8 months ago) |
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Quote:
rorschachX said:
Quote:
lsatrap said:
I still think it looks as if the mold spores where inoculated into the substrate.It doesn't look as if it was already in the sub and then grew. It looks like it was injected in.It might not be the vendors fault but I think the contams where introduced through inoculation.
Absolutely could be. Easy test would to set aside a grain jar after sterilizing it and don't inoculate it. It would just allow you to say for certain it is the spore syringe.
My FOAF always sets aside 1 grain jar each time he knocks up a set of jars just for this reason.
Great idea.
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