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 rorschachX
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Registered: 08/16/09
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Last seen: 14 years, 6 months
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 Advice needed; high contam rate recently. 
#10872831 - 08/16/09 09:19 PM (14 years, 7 months ago) |
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My friends friend was going to put this originally in the contamination forum but I thought about it and my friends friend knows he's contam'ing and what kind it is, instead he needs advice as far as the next step to take.
Here is the information of what's happening and what my friends friend has done to try to counter it.
AFOAF originally started with PF tek like most people and this was easy. He even cut them in half and cased with good results. Then he moved on to grains. He chose white popcorn and had moderate success here; probably getting about 60% success rate using White popcorn and spawning to coir in mini tubs.
He decided to step up yet again to WBS to try to get the contam rate down and for the smaller grain to get more inoculation points when spawning to bulk. He got WBS that had a high cracked corn content. Dreading the contam rate he got with popcorn he used a colander to sift out most of the cracked corn and let the milo and millet fall through. He then proceeded to wash and remove the floaters. He washed it very well reaching 100% clarity when the WBS was stirred in a 5 gallon bucket and no floater (he said it apparently took anywhere from 20-30 washes of 5 gallons of water to reach this level of cleanliness.). He then simmered for 15 minutes; dumped and allowed to dry. He often found that allowing it to air dry resulted in the grain being too wet so he applied a towel and a hair dryer set on a low setting to help achieve an even dryness. The inside of the grain was still saturated but the outside of the grain was dry to the touch. He then put the warm seeds into quart jars and allowed to sit for 20 minutes at which point he would dump the seeds out into a bowl to further remove the condensation that formed inside the jars. If he did not do this he found that after PCing the jars the moisture content of the jars looked to be too much.
After the above was completed he would PC for 90 minutes. He previously did 60 minutes and had high contam rates. Tiring of having all his efforts wasted he opted for 90 minutes.
He then used some liquid culture that he made previously by taking a single fully colonized popcorn kernel and putting it into a Karo mix. This LC is very well colonized; better than he thought would happen on his first LC attempt.
The results of every WBS jar done this way has resulted in black pin mold.
He then decided that it was an environment issue and sterilized one of his bathrooms. He washed the walls; the mirror, counters, floor, doors, trim, EVERYTHING with a 15% bleach solution. He then went into the attic and removed the ducts from the vents to prevent air from moving into this room. He also sealed the doors to this room. In addition to this he started showering before working with the jars; put on a full tyvek suit with goggles, dust mask, latex gloves, sterile booties and worked inside of a glove box with saran wrap curtains.
He still got black pin mold.
A few things to note:
He uses tyvek filters made from FedEx envelopes on the quart mason jars.
He shoots 3cc of LC into each jar; covering each inoculation hole with a band-aid.
With the popcorn he was originally using an incubation chamber (tub in tub) but because of the 40% contam rate he decided to leave the WBS jars at room temperature inside the glove box which had been sterilized and sprayed with Oust with a saran wrap curtain. Contam rate with WBS has been 100%.
Prior to moving to WBS he had tried yellow popcorn instead of white popcorn and also had 100% contamination rate; however a grain to grain transfer allowed 60-70% colonization before black pin mold reared it's ugly head. He used about 3-4 kernels to do this grain to grain per quart jar.
He is currently using a 6qt presto pressure cooker; feeling uncomfortable with the quality and questioning whether he was reaching the proper pressure he decided to buy an All American 1925x Stove top Autoclave. It was a great deal. Brand new on ebay for $99 instead of the retail cost of approx $300. It has not arrived yet but he is hoping that perhaps this is his problem.
He also questions whether this particular brand of WBS is just crap; however it would stand to reason that 90 minutes of pressure cooking time should be enough to kill the germinated spores.
He questions whether the LC being used is contaminated however looking at the liquid culture it looks very clean and healthy. Lots of thick mycelium growing and no indication of mold growth anywhere; not even on the popcorn kernel that is in it. The LC mix has 10% hydrogen peroxide mixed in when it was made.
The syringes he uses to draw the LC is sterilized in boiling water and then washed with 1CC of rubbing alcohol inside it. Tip is flame sterilized and alcohol wiped prior to injecting into the jars.
My friends friend at this point is just looking for advice on what additional steps he should take and in what order. Should he switch grains; perhaps back to white popcorn and see if he still has 100% contam rate? Should he wait for his new AA 1925x to see if the crappy walmart Presto PC is the problem? Or should he toss his LC and order new spores (this is the only stuff he has to inoculate with; no spore syringes from vendors with him. Only the LC syringes he made)?
Would trying to grow the LC using the cardboard tek be recommended to see if black pin grows on it as well?
He feels fairly confident that he knows what he's doing since he had some success originally.
I know this is long winded but I wanted to be detailed since I'm posting this for my friends friend. I appreciate anyone who reads this and any advice they can give. He feels he's on the right path to eliminate other things from being the source of contamination but wanted to see if anyone else saw flaws in his methods used thus far and if they also felt he was in the currently going in the right direction to correct this and in what order he should do them in.
Thank you very much.. my friend has spent weeks and tons of wasted time/effort/work/reading the forums before deciding to ask me to post this on here.
Again Thank you,
-Rorschach
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rorschachX
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Registered: 08/16/09
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Last seen: 14 years, 6 months
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10872862 - 08/16/09 09:24 PM (14 years, 7 months ago) |
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Just incase you need a mental image of how sterile this guy looks/is.. When he is working in that sterile bathroom he looks like he is one of the clean up guys from hazmat. People make fun of him saying he looks ridiculous and he's being paranoid/overkill; but my friends friend doesn't care better to be safe than sorry he says.
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RogerRabbit
Bans for Pleasure
 
Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 1 month
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Re: Advice needed; high contam rate recently. (moved) [Re: rorschachX]
#10872866 - 08/16/09 09:24 PM (14 years, 7 months ago) |
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 This thread was moved from Mushroom Cultivation.
Reason:
Your friend's friend should have stuck with his original idea and put it in the correct forum.
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rorschachX
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Last seen: 14 years, 6 months
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Re: Advice needed; high contam rate recently. (moved) [Re: RogerRabbit]
#10872878 - 08/16/09 09:25 PM (14 years, 7 months ago) |
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My apologies. Thank you for moving the thread.
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 Doc_T
Random Dude
Registered: 03/06/09
Posts: 42,395
Loc: Colorado 
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10872882 - 08/16/09 09:26 PM (14 years, 7 months ago) |
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Hold onto the LC, it may or may not be to blame.
Get grain without cracked corn. Washing your grain 30-50 times is beyond stupid.
Small PC may or may not get to 15 or 12 or even 10 psi, no matter what brand. So wait until the AA rrives before yo run any more jars.
And read the PC instructions, make sure you are doing it right. Sounds obvious, I know, but you asked.
So, new PC, new grain, try the LC in a few jars, go from there.
Meanwhile, it sure wouldn't hurt to order more spores or trade for some.
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You make it all possible. Doesn't it feel good?
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rorschachX
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Re: Advice needed; high contam rate recently. [Re: Doc_T]
#10872921 - 08/16/09 09:33 PM (14 years, 7 months ago) |
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Quote:
Doc_T said:
Hold onto the LC, it may or may not be to blame.
Get grain without cracked corn. Washing your grain 30-50 times is beyond stupid.
Small PC may or may not get to 15 or 12 or even 10 psi, no matter what brand. So wait until the AA rrives before yo run any more jars.
And read the PC instructions, make sure you are doing it right. Sounds obvious, I know, but you asked.
So, new PC, new grain, try the LC in a few jars, go from there.
Meanwhile, it sure wouldn't hurt to order more spores or trade for some.
Why is washing 30-50 times stupid. The water does not go to waste since it is used for the garden in his back yard. In addition there are many floaters and apparently lots of starch (water stays very murky up into the 20-25 washes with enough seed for 4-5 jars). Starch is known to be a breeding ground for molds. I'm not arguing but rather trying to understand why this shouldn't be done. It was my friends understanding from reading here that you want to clean it as best as possible.
For the PC he was leaving the weight off the pressure cooker and waiting to see a steady stream of steam coming out of the PC before placing the weight on it to be sure that all the current air occupying it had been evacuated. Pretty sure this is correct for any PC.
Thanks for your input even if you think it's obvious. Sometimes it is the obvious things that get missed.
And my friend's friend will definitely take your advice on getting more spores. He had been eyeing the PE6 from Ralphsters anyway. 
Edited by rorschachX (08/16/09 09:37 PM)
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Doc_T
Random Dude
Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10872939 - 08/16/09 09:37 PM (14 years, 7 months ago) |
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You want the grain clean. Wasting the time, if not the water, is stupid. Unless you just like stirring stuff around in water I mean.
Otherwise get some cleaner grain. Penningtons' is a favorite brand of WBS, but rye and other grains are cleaner and are easier to work with too.
PC handling right up to that point- then you need to turn it down as low as it can go and maintain pressure. That's when you start the timer.
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You make it all possible. Doesn't it feel good?
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rorschachX
Stranger
Registered: 08/16/09
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Last seen: 14 years, 6 months
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Re: Advice needed; high contam rate recently. [Re: Doc_T]
#10872963 - 08/16/09 09:40 PM (14 years, 7 months ago) |
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Quote:
Doc_T said:
You want the grain clean. Wasting the time, if not the water, is stupid. Unless you just like stirring stuff around in water I mean.
Otherwise get some cleaner grain. Penningtons' is a favorite brand of WBS, but rye and other grains are cleaner and are easier to work with too.
PC handling right up to that point- then you need to turn it down as low as it can go and maintain pressure. That's when you start the timer.
Indeed he does hate the time wasted. Looked for pennington but the local stores here seem to not have it. He will check some feed stores or Whole Foods for some Rye grain/berries; it seems that most people have better luck with this.
Correct; he turns it down until there is a very gentle rock and starts the timer.
Thank you for your advice; it is very much appreciated if for nothing else just to get some affirmation that he's on the right track.
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Triptonic
Registered: 04/23/08
Posts: 3,707
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10872992 - 08/16/09 09:44 PM (14 years, 7 months ago) |
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 I use this brand of WBS it has cracked corn in it...I dont see anything wrong with cracked corn. Wash your grains as much as you want. What I do is put them in a big pan and pour water on them, stir it up with my hand, then pour out the water. I repeat this step until the water is clear. Then I soak and then I simmer.
 These are the results I get after 3 days....no contams.
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Doc_T
Random Dude
Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10872996 - 08/16/09 09:46 PM (14 years, 7 months ago) |
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Yep, sounds like the right track. Narrow it down one thing at a time.
Keep an eye on the jars where the LC seemed to work, look for subtle signs that something is wrong.
Could be your local spore load is so high that you are just doomed and you need to take a few months off. Clean it all up, put it away, and just coast on the fruits you have. Then fire it up next spring with a new outlook and new knowledge.
I didn't read all of that, are you using a glovebox?
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You make it all possible. Doesn't it feel good?
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rorschachX
Stranger
Registered: 08/16/09
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Re: Advice needed; high contam rate recently. [Re: Doc_T]
#10873041 - 08/16/09 09:55 PM (14 years, 7 months ago) |
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Quote:
Doc_T said:
Yep, sounds like the right track. Narrow it down one thing at a time.
Keep an eye on the jars where the LC seemed to work, look for subtle signs that something is wrong.
Could be your local spore load is so high that you are just doomed and you need to take a few months off. Clean it all up, put it away, and just coast on the fruits you have. Then fire it up next spring with a new outlook and new knowledge.
I didn't read all of that, are you using a glovebox?
Yes he is using a glove box with a saran wrap curtain that has been sterilized with bleach water and sprayed with oust with all his tools inside it.
My friends friend also take a shower before he does his work; he puts on a tyvek jump suit with the hoodie, latex gloves, sterile booties a dust mask and goggles.
A/c has been disconnected from that room. There is no carpet in there. And the entire room has been sterilized with bleach water (every inch of it.. ceiling to floor.).
My friend has also debating covering the entire area in panda film but he doubts this would provide any more of a sterile environment than just using bleach water to wipe it down floor to ceiling and using a glove box.
And to triptonic; due to the amount of endospores that he's read cracked corn contains and the 40% contamination rate he had with popcorn he is trying to avoid anything containing corn if possible. The oils and starches from the cracked corn seem to accelerate the pin mold growth from his observations when compared to the previous jars he did have success with. When he filtered out 90% of the cracked corn pin mold growth occured but at a much delayed rate; probably as much as 5-6 days later compared to jars that did not have cracked corn filtered out.
He believes it's the WBS but due to several factors that are unknowns (PC, grain, liquid culture) he didn't want to lay definitive blame on the WBS.
Edited by rorschachX (08/16/09 10:01 PM)
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rorschachX
Stranger
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10873379 - 08/16/09 11:02 PM (14 years, 7 months ago) |
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The city in which we live has listed the spore count here as Low as of Sunday, August 16th. According to the city website Low means 1-6499 per cubic meter of air.
Not sure if that helps but it sounds to me like I should be in the clear as far as excessive mold spore count being the culprit.
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Doc_T
Random Dude
Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10874498 - 08/17/09 06:16 AM (14 years, 7 months ago) |
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Quote:
rorschachX said:
The city in which we live has listed the spore count here as Low as of Sunday, August 16th. According to the city website Low means 1-6499 per cubic meter of air.
Not sure if that helps but it sounds to me like I should be in the clear as far as excessive mold spore count being the culprit.
That's the city's count of allergenic spores. Not contam spore rate in your apartment.
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You make it all possible. Doesn't it feel good?
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Grumble121
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10877085 - 08/17/09 04:56 PM (14 years, 7 months ago) |
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Heh, I have really good luck with contams, with cheap bird seed with cracked corn. I cook the hell out of my bird seed; long story, but it is very hard to get my PC up to 15 PSI.
It usually ends up spending many hours at high temperature, but not 15 PSI. I try to be careful with sterile technique, but I must admit I can be fairly sloppy.
I don't always flame needles, I do not wipe the needle with alcohol between jars, sometimes I neglect to wear a mask, and oftentimes I am not careful about touching things outside my "glovebox"(a garbage bag, with lots of lysol), such as jars sitting out in the open, awaiting inoculation.
Whatever I am doing, it works; 0 contams in about 60 jars, 30 of which were grain to grain transfers using a garbage bag as a glove box.
I seem to recall the feed store mentioning something about grain being pre-sterilized. I almost am tempted to try a boil only jar, just to see what would happen.
Conventional wisdom says it would contam like crazy, but I am not so sure anymore with the stuff I use.
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rorschachX
Stranger
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Re: Advice needed; high contam rate recently. [Re: Grumble121]
#10877403 - 08/17/09 05:49 PM (14 years, 7 months ago) |
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Quote:
Grumble121 said:
Heh, I have really good luck with contams, with cheap bird seed with cracked corn. I cook the hell out of my bird seed; long story, but it is very hard to get my PC up to 15 PSI.
It usually ends up spending many hours at high temperature, but not 15 PSI. I try to be careful with sterile technique, but I must admit I can be fairly sloppy.
I don't always flame needles, I do not wipe the needle with alcohol between jars, sometimes I neglect to wear a mask, and oftentimes I am not careful about touching things outside my "glovebox"(a garbage bag, with lots of lysol), such as jars sitting out in the open, awaiting inoculation.
Whatever I am doing, it works; 0 contams in about 60 jars, 30 of which were grain to grain transfers using a garbage bag as a glove box.
I seem to recall the feed store mentioning something about grain being pre-sterilized. I almost am tempted to try a boil only jar, just to see what would happen.
Conventional wisdom says it would contam like crazy, but I am not so sure anymore with the stuff I use.
Not to be rude but what was the point of posting this besides advocating bad practices? Can't say my friends friend would take advice you have to give especially since he knows his problem is 1 of 3 things and he is very sterile.
Good for you that you've gotten away with such sloppy work; not all of us can be so lucky. The city we live in is riddled with allergens, pollen and spore count typically. It's very warm and humid here all year round.
This wasn't meant to be rude but more so to point out the ridiculousness of that post. I guess thanks for your input though.
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rorschachX
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10877428 - 08/17/09 05:53 PM (14 years, 7 months ago) |
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Let me follow up on my last post so it's clear the message I was trying to get across for my friend.
It's great that you've been so lucky thus far and had no contamination rate; but I wouldn't be boasting about it. There is a right and wrong way to do things.
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rorschachX
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10877938 - 08/17/09 07:25 PM (14 years, 7 months ago) |
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My friend will have me update this thread again once he has determined the cause and will outline the steps taken to remedy the solution; if for nothing else to help other people when they use the search function.
Thank you Doc_T and Triptonic.
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rorschachX
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10910018 - 08/22/09 04:50 AM (14 years, 7 months ago) |
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Update:
Got the All American 1925x stove top sterilizer. My friend's friend got all his jars ready.. pre sterilized them just to get used to using his new toy. He then went to check on his LC and there was what he believed to be black mold.
I am willing to bet this was the culprit; however my friend is not ready to place blame yet. He has syringes on order and will save 2cc's to make a new LC from spores as opposed to trying to make it from a grain of colonized popcorn.
He is fairly confident that he could probably go back to popcorn now with his new Sterilizer. He said it was much easier to use than the crappy presto pressure cooker and doesn't know how he got around without an AA pressure cooker/sterilizer before. It really is of the highest quality. The best part for him is that he can fit 8 jars in there (6 jars in the normal position and 2 jars on top of them on their side with plenty of room to spare). Coming from doing 2 jars at a time this is a huge time saver.
Will update once again once my friend get's his spores and has a successful inoculation; again just for search purposes for others who might have the same frustration my friend had.
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Acidium
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10930814 - 08/24/09 11:23 PM (14 years, 7 months ago) |
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Ive been reading this thread today, I havent had enough expertise as far as growing yet to give any advice or I wouldve. I just wanted to throw in that I hope all is well with the new batch your friend makes. Im willing to bet the LC was the culprit as well.
Let us know what the results are!
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rorschachX
Stranger
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Re: Advice needed; high contam rate recently. [Re: Acidium]
#10944646 - 08/26/09 09:12 PM (14 years, 7 months ago) |
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Quote:
Acidium said:
Ive been reading this thread today, I havent had enough expertise as far as growing yet to give any advice or I wouldve. I just wanted to throw in that I hope all is well with the new batch your friend makes. Im willing to bet the LC was the culprit as well.
Let us know what the results are!
Thanks for the well wishes. My friend appreciates the thoughts. He is confident that it was his LC; although he's still unsure if his PC was ever hitting 15psi. In either case he has thrown out the LC, has spores that should be arriving tomorrow, and has an All American 1925x sterilizer that can fit 8 jars. He feels that with these 2 things now being taken out of the picture he should start getting some uncontam'd jars. He is very excited about this since he has just set up a martha; an upgrade from monotubs. He likes that you have more environmental control in a martha than in other setups.
A New LC jar will be started tomorrow as well as the grains will start soaking. He found a good source of bird seed that is about 85% millet; he also bought 2 bags of popcorn. He's going to do a half the jars with WBS the the other half with popcorn. He's read countless threads on the topic but he thought now that he has the proper tools, spores and sterile environment that he would just experience both grains for himself to decide.
Thanks again.
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Doc_T
Random Dude
Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#10962267 - 08/29/09 04:48 PM (14 years, 7 months ago) |
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Instead of starting the LC from spores, use Grain LC from a colonized jar,
That will help a bunch.
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You make it all possible. Doesn't it feel good?
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rorschachX
Stranger
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Last seen: 14 years, 6 months
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Re: Advice needed; high contam rate recently. [Re: Doc_T]
#11048851 - 09/12/09 09:52 PM (14 years, 7 months ago) |
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Update as promised:
My friend of a friend seems to be having luck. Out of 18 jars only 1 shows signs of potential problems.
AFOAF has done 6 jars of popcorn with Malabar Coast, 6 jars WBS of Mazatapec, 6 jars of rye with Penis Envy.
So far the Popcorn appears to be going the fastest however for spawning to bulk the WBS seems like a better choice although it seems slow. My FOAF didn't do the rye until like 5-6 days later and it's now showing very good growth and could indeed actually be the fastest colonizer. It is hard to say because they are all different "strains", some might be more aggressive at colonizing than others.
My FOAF's liquid culture is also looking good and once he spawns these jars he will start up a new batch and knock them up with PE LC.
One Jar seems to have some green mold growing in a very small area but the rest of the jar is growing mycellium at a very fast rate. My FOAF is going to let this grow out in the jar but most likely toss it. He wants to see how Rye does and if the Mycellium will out grow the mold.
He is planning on spawning 2 quarts grain per 6.5quart storage container. I guess this is a 1:3 ratio?
In any case the AA 1925x seems to have fixed most of my friends problems; in addition he now has a known good/sterile LC to work from.
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rorschachX
Stranger
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Last seen: 14 years, 6 months
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#11048890 - 09/12/09 10:02 PM (14 years, 7 months ago) |
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So my FOAF did 18 jars and only 1 contam'd.. the rest are looking great. That's a 94% success rate (I think). That's not bad, going from pretty much 100% failure to 94% success; looks like he got his procedure and proper equipment together.
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rorschachX
Stranger
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#11134095 - 09/26/09 09:12 PM (14 years, 6 months ago) |
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Question.
My FOAF had 1 rye jar that got infected early on with what looked like trich. He set it aside from the other jars but let it grow out. The mycellium has now taken over the trich and it is a very bright white color. Does this mean the Mycellium has consumed the trich??? Or is the jar still pretty much contamed?
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Triptonic
Registered: 04/23/08
Posts: 3,707
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Re: Advice needed; high contam rate recently. [Re: rorschachX]
#11134111 - 09/26/09 09:15 PM (14 years, 6 months ago) |
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Shake the jar and you will find out I bet its just dormant in the center of the jar. I would throw it out. But I'm almost 100% sure that after you shake it you will find out that it is still infected. Unless it never was in the first place. SHAKE THAT BITCH!
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